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PubMedDate not supplied

First isolation of bacteriophages for a spirochaete: potential genetic tools for Leptospira.

Three bacteriophages of the saprophytic aquicole bacterium Leptospira biflexa were isolated from sewage waters from the outskirts of Paris, France. These phages do not infect representative strains of the pathogenic species Leptospira interrogans, and their host range is restricted to serovar patoc of the saprophytic species. The phages were found to be lytic and no lysogenic state could be demonstrated. Electron micrographs showed that the phages were morphologically identical and had polyhedral heads and contractile tails. Their genomes were sensitive to restriction enzymes and consisted of double-stranded DNA. Pulsed-field agarose gel electrophoresis indicated that their genomes were linear: 60 kb for LE1 and 50 kb for LE3 and LE4.

Bacteriophages↗
PubMed2005

Whole-genome analysis of Leptospira interrogans to identify potential vaccine candidates against leptospirosis.

Leptospirosis is an important global human and veterinary health problem. Humans can be infected by exposure to chronically infected animals and their environment. An important focus of the current leptospiral research is the identification of outer membrane proteins (OMPs). Due to their location, leptospiral OMPs are likely to be relevant in host-pathogen interactions, hence their potential ability to stimulate heterologous immunity. The existing whole-genome sequence of Leptospira interrogans serovar Copenhageni offers a unique opportunity to search for cell surface proteins. Predicted genes encoding potential surface proteins were amplified from genomic DNA by PCR methodology and cloned into an Escherichia coli expression system. The partially purified recombinant proteins were probed by Western blotting with sera from human patients diagnosed with leptospirosis. Sixteen proteins, out of a hundred tested, were recognized by antibodies present in human sera. Four of these proteins were conserved among eight serovars of L. interrogans and absent in the non-pathogenic Leptospira biflexa. These proteins might be useful for the diagnosis of the disease as well as potential vaccine candidates.

Antigens, Bacterial↗
PubMed2005

Characterization of IS1501 mutants of Leptospira interrogans serovar pomona.

Leptospira interrogans is a diverse species in which individual serovars have distinctive restriction fragment length polymorphisms that are useful in strain identification. Many of these polymorphisms can be detected using hybridization probes derived from insertion sequences; an observation that suggests these IS elements are active and can transpose in L. interrogans. Two spontaneous mutants of L. interrogans serovar Pomona strain RZ11 were isolated by immune selection and characterized. Changes in the size and antigenicity of LPS from these mutants were detected. Genetic analysis showed that both mutants have additional copies of an IS3-like element, designated IS1501, that are not present in the parental strain. One mutant, GT211, has a single additional copy of IS1501, whereas the other mutant, GT210 has three additional copies of IS1501 relative to strain RZ11. IS1501 transposition generated 3-bp direct repeats from target sequences flanking the insertion site. RT-PCR analysis of transcripts at altered loci showed IS1501 transcripts extended into adjacent sequences. These data are the first to show spontaneous transposition of an endogenous Leptospira insertion sequence, and suggest that IS1501 may be capable of gene activation.

Antigens, Bacterial↗
PubMed2005

Buenos Aires, a new Leptospira serovar of serogroup Djasiman, isolated from an aborted dog fetus in Argentina.

This study describes the isolation of a new leptospiral serovar from the Djasiman group from an Argentinean aborted fetus of a dog. The strain was isolated from a culture of mixed liver and kidney tissue from one aborted dog fetus. Bitch's serum showed a titre of 1:800 against the new serovar and 1:400 or less against other serovars tested. Microscopic Agglutination test (MAT) with a panel of 38 rabbit anti-Leptospira sera representative for all pathogenic serogroups was performed to establish the putative serogroup of the isolated strain. Serovar identification was done by Cross-Agglutination Absorption Test (CAAT). Species determination was carried out on basis of sequence identity of primer pair G1/G2 generated PCR products from the isolate with those from reference strains belonging to the various pathogenic Leptospira species. Results showed that the new strain named, Baires, belongs to species L. interrogans sensu stricto and the serogroup Djasiman. However, the isolate could not be identified as any of the known serovars within this serogroup. These results allow us to conclude that the strain Baires represents a new serovar (Buenos Aires) of Djasiman serogroup.

Aborted Fetus↗
PubMed2006

Isolation of Leptospira noguchii from sheep.

The main goal of this study was to obtain new isolates of Leptospira spp. from sheep. A total of 10 kidney samples and 44 blood samples were collected from sheep slaughtered in Pelotas, Southern Brazil. One isolate was obtained which was identified by 16S rRNA gene sequencing and serogrouping to be Leptospira noguchii serogroup Autumnalis. Microscopic agglutination test (MAT) evaluation revealed that 4.5% of the sheep sera reacted against the Autumnalis serogroup. This is the first report of isolation of L. noguchii from sheep. Together these findings indicate that L. noguchii infections may be a potentially important veterinary problem in this domestic animal species.

Agglutination Tests↗
PubMed2002

The impact of Leptospira seropositivity on reproductive performance in sows in southern Viet Nam.

A serologic survey was conducted among sows in the Mekong delta in southern Viet Nam to investigate associations between leptospiral seropositivity and reproductive performance. Data were collected from a total of 339 sows in lactation or gestation, from four large-scale state farms on three occasions. The seroprevalence for Leptospira interrogans serovar (sv) autumnalis was 32%, for L. interrogans sv bratislava 29%, for L. kirschneri sv grippotyphosa 13%, for L. interrogans sv icterohaemorrhagiae 27%, for L. interrogans sv pomona 5%, and for L. borgpetersenii sv tarassovi 13%. The reproductive parameters number of days from weaning to service (WSI), number of piglets born, number of piglets born dead, and number of piglets born weak, were evaluated. Seropositivity for sv tarassovi was associated with 0.8 more dead piglets per litter (P = 0.06), and sv grippotyphosa with a 1 day longer WSI (P = 0.06). There were no significant associations between reproductive performance and sv autumnalis, sv bratislava, sv pomona, and sv icterohaemorrhagiae. It is concluded that seropositivity for Leptospira can be associated with impaired reproductive performance even in areas where a high degree of immunity among sows is expected.

Animal Husbandry↗
PubMed1999

A DNA fragment of Leptospira interrogans encodes a protein which shares epitopes with equine cornea.

Horses infected with Leptospira interrogans present several clinical disorders, one of them being recurrent uveitis. An antigenic relationship between this bacterium and equine cornea has been described in previous studies. With the aim to make progress on defining the molecular basis and pathogenesis of equine recurrent uveitis, here we describe the cloning of one DNA fragment from a Leptospira interrogans serovar pomona genomic lambda gt11 library. Although there are references of transcription of leptospiral genes in E. coli from their own leptospiral promoters, in this recombinant construction the leptospiral DNA was located under the control of lacZ promoter since no expression could be detected in the absence of IPTG. This clone, isolated by expression screening with polyclonal serum raised against equine corneal proteins, encodes a 90 kDa protein of L. interrogans which crossreacts with equine cornea as proved Western-blotting. Antibodies directed against this leptospiral protein strongly recognised a 66 kDa equine corneal protein, one of those recognised by an anti-equine cornea serum. Our findings suggest that an immune response to 90 kDa protein participates in pathogenesis of equine uveitis.

Animals↗
PubMed1997

Leptospira interrogans serovar icterohaemorrhagiae seropositivity and the reproductive performance of sows.

The reproductive performance of 28 sows seropositive to Leptospira interrogans serovar icterohaemorrhagiae was compared with that of 87 Leptospira sp. seronegative dams belonging to the same herd. Sows were sampled during 1988 to 1993. During this period the herd was not submitted to any kind of intervention (antibiotic therapy, immunoprophylaxis or rodent control). Relative risks (RR) of return to heat, mummified fetuses, stillbirth, and weak newborn piglets for infected sows were assessed and the differences in means of total piglets born per litter, piglets born alive, piglets effectively housed, weaned piglets, stillbirths, mummified fetuses, weak newborn piglets, weight at birth of the piglets effectively housed, weight at 21 days of life and weight at weaning were evaluated. Seropositive dams had a greater risk of having weak newborn piglets (RR = 1.67, 1.02 < or = CI 95% < or = 2.72) and also of having more weak newborn piglets per litter (P = 0.01). Other variables examined were not different (P > 0.05).

Aging↗
PubMed1999

Identification of Leptospira biflexa by real-time homogeneous detection of rapid cycle PCR product.

Sequence analysis of 16S rRNA genes extracted from nucleic acids databases enabled the identification of a Leptospira biflexa (L. biflexa) signature sequence, against which a reverse primer designated L613, was designed. This primer, when used in conjunction with a universal bacterial specific forward primer designated Fd1, enabled the development of a LightCycler-based PCR protocol in which fluorescence emission due to binding of SYBR Green I dye to amplified products could be detected and monitored. A melting temperature (Tm), determined from the melting curve of the amplified product immediately following the termination of thermal cycling, confirmed that the product was that of L. biflexa. Agarose gel electrophoresis therefore was not necessary for identification of PCR products. The PCR protocol was very rapid, and consisted of 30 cycles with a duration of 20 s for each cycle with the monitoring of the melting curve requiring an additional 3 min. The whole protocol was completed in less than 20 min. The PCR protocol was also specific and enabled the identification of 18 strains of L. biflexa, whilst excluding 14 strains of L. interrogans and Leptonema illini. Two examples of its utility in improving work flow of a Leptospira reference laboratory are presented in this article. The use of a simple boiling method for extraction of DNA from all the members of the Leptospiraceae family DNA further simplifies the procedure and makes its use conducive to diagnostic laboratories.

Base Sequence↗
PubMed2003

Comparison of three different leptospiral vaccines for induction of a type 1 immune response to Leptospira borgpetersenii serovar Hardjo.

Leptospira serovar Hardjo are bacterial pathogens of cattle that cause zoonotic infections of humans. Monovalent serovar Hardjo vaccines protect cattle from serovar Hardjo while pentavalent vaccines do not even though they contain serovar Hardjo organisms. Here, cattle vaccinated with either of two monovalent vaccines had lymphocytes that made interferon-gamma (IFN-gamma) and IgG(1) and IgG(2) antibodies to Hardjo antigen while those from cattle vaccinated with a pentavalent Leptospira vaccine did not. IFN-gamma-producing cells were mainly CD4(+), but included CD8(+) and gamma delta TCR(+) cells. Despite their monovalent composition, those vaccines also induced IFN-gamma responses to serovar Grippotyphosa antigens. Thus, induction of a type 1 immune response is consistent with protective immunity to serovar Hardjo infections.

Animals↗
PubMed2000

Interest of partial 16S rDNA gene sequences to resolve heterogeneities between Leptospira collections: application to L. meyeri.

This paper describes the advantage of using the first 330 bp (positions 46 to 375, Escherichia coli numbering) of the 16S rDNA gene for comparison of Leptospira isolates. Phylogenetic analysis conducted from the whole 16S rDNA sequences available in databanks as well as that conducted from the partial sequences yielded quite similar results, in accordance with data inferred from previous DNA-DNA relatedness studies. This tool was used for the comparison of Leptospira strains from different reference collections. Consistent results were obtained from the analysis of the polymorphism generated by pulsed-field gel electrophoresis. The study focused on different serovars of L. meyeri species, the classification of which has been controversial. The results revealed large collection heterogeneities, and suggest that the classification of the L. meyeri species should be revised.

Bacterial Typing Techniques↗
PubMed1990

Immunological reactivity and passive protective activity of monoclonal antibodies against protective antigen (PAg) of Leptospira interrogans serovar lai.

Monoclonal antibody (MAb) AG1 against the protective antigen (PAg) was produced and characterized. It had been extracted from Leptospira interrogans serovar lai by the chloroform-methanol-water method and was of glycolipid nature (23-30Kd). The fact that the PAg was a serogroup-specific antigen was shown by MAb AG1, because MAb AG1 agglutinated serovars of serogroup Icterohaemorrhagiae. Purified MAb LW2 and LW3 which are agglutinating antibodies of serovar lai and AG1 passively protected hamsters from leptospiral infection. Induction of the reactive oxygen intermediates by MAbs from peritoneal exudate macrophages of mice were observed in the chemical luminescence assay and the MAbs reacted with the PAg in the dot enzyme-linked immunosorbent assay. However, MAb LW4a against the genus-specific antigen present in the sub-surface of leptospiral cells did not show protective and reactive-oxygen-inducing activities; they reacted with the non-protective glycolipid antigen of low molecular weight (Fr I, 10-15Kd) in the dot enzyme-linked immunosorbent assay. These results indicated that anti-PAg antibody exhibited opsonic activity against Leptospira and the production of reactive oxygens by macrophages led to leptospiricidal action as one of the defence mechanisms of the host against leptospiral infection. However, the antibody against the genus-specific glycolipid antigen may not be important for protection against leptospiral infection.

Agglutination Tests↗
PubMed1985

The coypu as a rodent reservoir of leptospira infection in Great Britain.

The coypu (Myocastor coypus Molina) is an aquatic rodent that has become a widespread pest in the south-east of England. Since the natural habitat of this animal is aquatic, the possibility of infection with leptospires was investigated. Twenty-nine coypu were trapped and examined by serological, histological and cultural methods. Of these, there was serological evidence of infection in seven coypus and Leptospira interrogans var. Wolffii was isolated from a further animal. This appears to be the first report of the isolation of leptospira from a coypu in Great Britain.

Animals↗
PubMed1981

The occurrence and significance to animal health of Leptospira, Mycobacterium, Escherichia coli, Brucella abortus and Bacillus anthracis in sewage and sewage sludges.

Samples of sewage, sewage sludge and sewage effluent from one or more of four sewage treatment plants were examined for the presence of Leptospira, Mycobacterium, Escherichia coli, Brucella abortus and Bacillus anthracis. Brucella abortus and Bacillus anthracis were not isolated. Eleven strains of E. coli potentially enteropathogenic for calves or piglets, eight pathogenic strains of Mycobacterium and one patho;genic Leptospira strain were isolated from 101, 189 and 189 samples respectively. Sewage sludge is no;t considered to play a major part in the epidemiology of disease caused by these organisms.

Animal Diseases↗
PubMed1995

Prevalence of Leptospira spp. in wild brown rats (Rattus norvegicus) on UK farms.

Wild brown rats (Rattus norvegicus) are frequently implicated in the carriage and spread of Leptospira spp. Wild brown rats (n = 259) were trapped from 11 UK farms and tested for Leptospira spp. using a number of diagnostic tests. The prevalence of leptospiral infection was low, but there was variation in the results obtained with the different diagnostic tests. Estimates of prevalence ranged between 0% by silver-staining of tissues, 1% by the microscopic agglutination test, 4% by the enzyme-linked immunosorbent assay, 4% by culture, and 8% by fluorescent antibody technique. In total, 37 (14%) rats were positive by at least one of the tests, which contrasts with the frequently reported prevalences of 50-70% for wild rats in the UK. Serovar bratislava was a prevalent as icterohaemorrhagiae, although it was present only on farms with larger rat populations.

Animals↗
PubMed1998

The prevalence of antibody to the viruses of bovine virus diarrhoea, bovine herpes virus 1, rift valley fever, ephemeral fever and bluetongue and to Leptospira sp in free-ranging wildlife in Zimbabwe.

The prevalence of antibody to the viruses of bovine virus diarrhoea (BVD), bovine herpes virus typel (BHV1), rift valley fever (RVF), bovine ephemeral fever (BEF) and bluetongue (BT) and to Leptospira sp. was determined in wildlife populations in Zimbabwe. Evidence of infection with BVD virus was found in 14 of the 16 species examined but was greatest in eland Taurotragus oryx, nyala Tragelaphus angasi and bushbuck Tragelaphus scriptus. Persistent infection with BVD virus was found in 1 of 303 antibody-free eland but not in the smaller sample of 102 antibody-free buffalo Syncerus caffer. Antibody to BHV1 was widespread, being found in 10 of 16 species with the highest prevalence being in buffalo and eland. Antibody to RVF was most prevalent in black rhino Diceros bicornis and white rhino Ceratotherium simum, buffalo and waterbuck Kobus ellipsiprymnus. Both BEF and BT were widespread in all the species examined. Evidence of infection with Leptospira sp. was found in 7 species. Infections were due to up to 3 of 8 different serovars.

Animals↗
PubMed2000

Prevalence of antibodies to Leptospira serovars in sheep and goats in Alto Adige-South Tyrol.

Serum samples from 313 sheep and 95 goats were collected during November 1993 in 26 localities in Alto Adige-South Tyrol and tested by microscopic agglutination test for antibodies to 28 serovars of the genus Leptospira. At the time of blood collection all the animals appeared healthy with no clinical sign suggestive of leptospirosis. The observed seroprevalence in sheep was 6.1%, whereas the seropositivity rate for goat serum samples was 2.1%. The highest serological prevalence in sheep was recorded for serovar castellonis, followed by poi, sejroe, hardjo subtype hardjobovis, copenhageni, and cynopteri. Titres to poi were the only ones found in goats. These findings, which are proof of Leptospira infection in Alto Adige-South Tyrol, indicate that foci of several serovars exist in this region.

Animals↗