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Environmental exposure and leptospirosis, Peru.

Human infection by leptospires has highly variable clinical manifestations, which range from subclinical infection to fulminant disease. We conducted a population-based, cross-sectional seroepidemiologic study in Peru to determine potential relationships of environmental context to human exposure to Leptospira and disease associated with seroconversion. Three areas were studied: a flooded, urban slum in the Peruvian Amazon city of Iquitos; rural, peri-Iquitos villages; and a desert shantytown near Lima. Seroprevalence in Belen was 28% (182/650); in rural areas, 17% (52/316); and in a desert shantytown, 0.7% (1/150). Leptospira-infected peridomestic rats were found in all locales. In Belen, 20 (12.4%) of 161 patients seroconverted between dry and wet seasons (an incidence rate of 288/1,000). Seroconversion was associated with history of febrile illness; severe leptospirosis was not seen. Human exposure to Leptospira in the Iquitos region is high, likely related both to the ubiquity of leptospires in the environment and human behavior conducive to transmission from infected zoonotic sources.

Adolescent↗

[Epidemiological aspects of leptospirosis in the eastern regions of the Russian Federation].

There have recently substantial changes in the basic parameters of Leptospira infection in the eastern regions. The infection is characterized by a pronounced and steady-state rise in morbidity rates, by its trend for urbanization, by the spread of the disease in recreational areas, by a primary role of dogs as sources of infection, by polymorphism and atypical nature of the clinical picture of Canicola leptospirosis, which make its diagnosis difficult. The anthropurgic foci where dogs and pigs are the main sources of pathogenic leptospiras are of the leading epidemiological significance. The optimum package of measures reducing a risk of human infection is recommended.

Adolescent↗

The efficacy of a leptospirosis vaccine in preventing leptospiruria in pigs.

A commercially manufactured leptospirosis vaccine containing serovars pomona and hardjo and licensed for use in cattle and sheep was investigated to determine if it would prevent leptospiruria in pigs exposed to serovar pomona. Twenty piglets were each vaccinated twice at an interval of three weeks. Twenty other piglets were unvaccinated and served as controls. Three weeks after the second dose of vaccine all animals were exposed for 64 to 89 days to a natural infection with pomona. During the investigation blood samples were examined serologically and urine samples were examined by dark ground microscopy and cultured for the presence of leptospirae. Attempts were made to culture leptospirae from kidneys at slaughter. Kidneys were also examined histologically for evidence of leptospira infection. One vaccinated animal developed a respiratory disease. It was treated with antibiotics and removed from the trial. Leptosphuria was demonstrated in six of the remaining 19 vaccinated pigs and leptospirae were found in nine of 578 (1.5%) urine samples examined from these animals during the period of exposure. In contrast leptospiruria occurred in 19 of 20 unvaccinated pigs and leptospirae were found in 253 of 642 (39.4%) urine samples examined from these animals. Histopathological lesions consistent with leptospirosis were found in kidneys examined from two of 16 vaccinates and 17 of 18 non-vaccinates. Antibodies to serovar pomona were detected in 12 of 19 vaccinated pigs examined three weeks after the second dose of vaccine and before exposure to infection, and in all of 18 unvaccinated pigs examined after exposure to infection. It was concluded that use of this vaccine in pigs resulted in a significant degree of protection against leptospiruria.

Journal Article↗

[Identification of infectious agents in cases suspicious of Hantavirus infection but with negative serology].

BACKGROUND: The diagnosis of acute respiratory illness caused by Hantavirus is based in the rapid and progressive clinical course, epidemiological background and the serological confirmation of the virus. When the presence of the virus is not confirmed a differential diagnosis must be made with other infections. Between 1999 and 2001, the Chilean Public Health Institute received 1063 blood samples from patients with a suspicious clinical picture, to study the presence of Andes strain of Hantavirus. In 134 of these samples, the presence of the virus was confirmed. AIM: To study the presence of other infections in sera from patients with suspected Hantavirus acute respiratory illness but serologically negative for Hantavirus. MATERIAL AND METHODS: A retrospective study of 98 serum samples, received at the National Public Health Institute, of patients with negative serology for Hantavirus. The presence of antibodies against influenza virus, Mycoplasma pneumoniae, Leptospira and Streptococcus pneumoniae was determined using Latex techniques. RESULTS: Leptospira was detected in 23 patients, influenza virus in 13, Streptococcus pneumoniae in six and Mycoplasma pneumoniae in one case. Leptospira infections were confirmed in four cases by ELISA determination of IgM antibodies. Influenza virus infection was confirmed in three cases by Hemmaglutation Inhibition Assay. CONCLUSIONS: In suspected cases of Hantavirus acute respiratory infection, but with negative serology, other infectious agents such as Leptospira, influenza virus and Streptococcus pneumoniae, must be sought.

Adolescent↗

Enzyme-linked immunosorbent assay for determining specific immunoglobulin M in infections caused by Leptospira interrogans serovar hardjo.

An automated enzyme-linked immunosorbent assay detecting specific immunoglobulin M in infections with Leptospira interrogans serovar hardjo was evaluated on 69 patients. The test was sensitive and simple to perform, requiring a single dilution of test serum, with data expressed as units of antibody activity interpolated from a reference serum pool.

Agglutination Tests↗

Epidemiology and characterization of leptospirosis at an urban and provincial site in Thailand.

Patients with FUOs at the Children's Hospital in Bangkok and the Chao Phya Abhai Bhu Bejhr Hospital in Prachinburi were screened for leptospirosis by blood and urine culture in addition to microagglutination testing of their serum. Animal populations in urban and periurban areas of Bangkok were surveyed for evidence of leptospira infection. Three rural sites near the Prachinburi Provincial Hospital were also surveyed. The rodents' and domestic animals' blood, urine, and/or kidney cell samples were cultured for leptospira. Sera from these animals were also tested for leptospira antibody. The bataviae serovar was the most commonly detected leptospiral agent in both man and animals. Presenting symptoms varied with age with children showing primarily fever, vomiting, headache, abdominal and generalized muscle pain and diarrhea whereas adults had fever, headache, anorexia, muscle pain and constipation. Blood samples from patients suspected of having leptospirosis were tested for antibody by the MAT and cultured in EMJH media. The following serogroups were identified: bataviae, autumanalis, javanica, hebdomadis, and pyrogens. Leptospirosis incidence in humans was much higher in the rainy/flooding year of 1983 compared to the relatively dry year of 1984. Results of our animal surveillance studies indicate that in addition to rats, which have previously been mentioned, dogs, bandicoots, cattle and pigs could be the source of human leptospirosis infection in both urban and provincial locations in Thailand.

Animals↗

Evaluation of a modified Taqman assay detecting pathogenic Leptospira spp. against culture and Leptospira-specific IgM enzyme-linked immunosorbent assay in a clinical environment.

A Taqman assay for the detection of pathogenic Leptospira was modified to suit the LightCycler instrument. The modified assay was found to have an analytical sensitivity of 10 copies/reaction. The assay was then compared to the current gold standard for acute phase detection, culture, and to a commercially available Leptospira-specific IgM enzyme-linked immunosorbent assay (ELISA). Of the 236 samples including serum and ethylenediaminetetraacetic acid anticoagulated blood sample submitted for testing, polymerase chain reaction (PCR) was able to detect Leptospira DNA in 27 of the 28 culture positives and in 1 negative culture. Discrepant results were resolved by using the microscopic agglutination test on convalescent sera. The PCR was found to have a clinical specificity and sensitivity of 99.5% and 96.4%, respectively, when compared to the culture. Comparisons between culture and ELISA showed that the ELISA lack sensitivity (4.2%) and positive predictability (3.6%) for the detection of acute phase Leptospira infections. These results show that the modified LightCycler Taqman assay could be used as a replacement of culture for the detection of pathogenic Leptospira in a clinical setting.

Antibodies, Bacterial↗

Development of species-specific PCR primer sets for the detection of Leptospira.

Spirochetes of the genus Leptospira infect animals and humans and are the causative agents for the emerging infectious disease leptospirosis. Rapid and simple assays for the identification of individual Leptospira species are currently not available. For identification of individual Leptospira species, PCR primers that detect the ompL1 gene sequence for the majority of pathogenic leptospires were developed in this study. The primer pairs detect Leptospira interrogans, Leptospira borgpetersenii, Leptospira kirschneri, Leptospira santarosai, Leptospira weilii and Leptospira noguchii, without cross-reacting with other Leptospira species. The development of the primers revealed a divergence of the ompL1 gene within L. interrogans, splitting this species into two separate groups. The species-specific primers will be especially useful in epidemiological studies and disease outbreak investigations for the detection of Leptospira species in human, animal and environmental samples.

Bacterial Outer Membrane Proteins↗

Leptospiral infection in aborted equine foetuses.

During an investigation of equine abortion, leptospiral infection was demonstrated in nine out of 22 foetuses examined by immunofluorescence and culture. Strains belonging to four serogroups (Australis, Pomona, Hebdomadis and Icterohaemorrhagiae) were isolated. The age of leptospira infected foetuses ranged from six months to term.

Abortion, Veterinary↗

[Comparison of 4 microscopy techniques for the diagnosis of leptospirosis in wild rodents in a rural area of Valdivia, Chile].

Kidneys of six different species of 93 wild rodents captured in the rural area of Valdivia (Chile) were simultaneously examined for leptospira infection by means of Levaditi silver stain, dark ground microscopy in wet smears, and immunofluorescence and immunoperoxidase techniques using pooled antiserum against hardjo and pomona serovars. Leptospira was shown to be present in 40 (43.0%) rodents. They were detected in five of the six species: Akodon olivaceus, Akodon longipilis, Rattus rattus, Oryzomis longicaudatus and Mus musculus. Levaditi's technique detected the highest number of positive samples (67.5%) and the dark field microscopy the lowest (32.5%). The detection of leptospira in the kidneys of these wild rodents suggests that their urine may play an important role in the dissemination of leptospirosis in this area of Valdivia.

Animals↗

Biological activity of a peptidoglycan extracted from Leptospira interrogans: in vitro studies.

Peptidoglycan (PG) has been isolated from some species of spirochaetes, including Leptospira interrogans. Although leptospiral PG has been chemically characterized, no study has been carried out on its potential biological activity. Since PG of Treponema and Borrelia is biologically active both in vivo and in vitro, we investigated the capacity of a leptospiral PG preparation to induce relevant biological effects. PG extracted from L. interrogans strain Teramo was mitogenic at 0.1 microgram ml-1 for human peripheral blood mononuclear cells (PBMC) since it increased the PBMC fraction positive for Ki-67, an antigen expressed by human proliferating cells; at 4 micrograms ml-1, PG was able to induce complement consumption and to stimulate leucocyte phagocytosis and the metabolic burst of resting as well as phagocytosing leucocytes. These findings indicate that Leptospira PG may play a role in modulating the immunocompetent cell functions and suggest that PG can contribute to the host response during Leptospira infection.

Cell Division↗

Evaluation of multivalent Leptospira fluorescent antibody conjugates for general diagnostic use.

Four lots of conjugate were evaluated for optimal dilution and degree of fluorescence produced with reference cultures and bovine and porcine leptospira isolates. One lot that uniformly produced better fluorescence was evaluated for sensitivity and specificity with reference cultures, isolates, culture-positive tissues, and 13 other bacterial species. Further evaluation of the conjugates was done with bovine, porcine, and ovine specimens submitted to a diagnostic laboratory. Leptospires were detected with the fluorescent antibody test (FAT) in 9 of 21 culture-positive bovine kidneys and were detected in diluted cultures when present at concentrations of 10(2)-10(3) organisms/ml. With the exception of Treponema hyodysenteriae, FAT's of other bacterial cultures produced minimal fluorescence or were negative. Positives were characterized by moderate to brilliant fluorescence of typical cell forms, and most nonspecific fluorescence was eliminated with a flazo-orange counterstain. The results indicated that the FAT utilizing multivalent conjugates could be used successfully as an additional method for diagnosis of leptospira infections.

Animals↗

Muskrats as carriers of pathogenic leptospires in The Netherlands.

Leptospires were isolated from 24 of 327 (7%) muskrats (Ondatra zibethicus) caught in The Netherlands. All isolates were identified as Leptospira interrogans. One isolate was typed as serovar copenhageni in the Icterohaemorrhagiae serogroup, one as serovar lora in the Australis serogroup. Twenty-one isolates showed a close relationship with serovars grippotyphosa, valbuzzi, muelleri and ratnapura from the Grippotyphosa serogroup. One isolate was lost. Sera from 196 muskrats were examined by the microscopic agglutination test. Forty-five (23%) sera reacted positively (titers greater than or equal to 1:160), 42 (21%) of these 45 sera to Grippotyphosa and 3 (2%) to Sejroe serogroup antigens. This is the first report of serological and cultural evidence of leptospira infection in muskrats in The Netherlands.

Animals↗

LruA and LruB, novel lipoproteins of pathogenic Leptospira interrogans associated with equine recurrent uveitis.

Recurrent uveitis as a sequela to Leptospira infection is the most common infectious cause of blindness and impaired vision of horses worldwide. Leptospiral proteins expressed during prolonged survival in the eyes of horses with lesions of chronic uveitis were identified by screening a phage library of Leptospira interrogans DNA fragments with eye fluids from uveitic horses. Inserts of reactive phages encoded several known leptospiral proteins and two novel putative lipoproteins, LruA and LruB. LruA was intrinsically labeled during incubation of L. interrogans in medium containing [14C]palmitic acid, confirming that it is a lipoprotein. lruA and lruB were detected by Southern blotting in infectious Leptospira interrogans but not in nonpathogenic Leptospira biflexa. Fractionation data from cultured Leptospira indicate that LruA and LruB are localized in the inner membrane. Uveitic eye fluids contained significantly higher levels of immunoglobulin A (IgA) and IgG specific for each protein than did companion sera, indicating strong local antibody responses. Moreover, LruA- and LruB-specific antisera reacted with equine ocular components, suggesting an immunopathogenic role in leptospiral uveitis.

Acylation↗

[Blood culture in the leptospiroses (author's transl)].

In the early stage of Leptospirae infection there is always blood stream invasion so blood culture is the best diagnostic test and permit precise etiologic identification. In the paper methods and selective media are suggested and the possibility of mistake so called "pseudo Spirochaetes" for Leptospirae is emphasized. Personal experience about the streght of blood culture in comparison with Martin and Pettit serumdiagnostic tests is reported.

Bacteriological Techniques↗

flaB-polymerase chain reaction (flaB-PCR) and its restriction fragment length polymorphism (RFLP) analysis are an efficient tool for detection and identification of Leptospira spp.

For establishment of a rapid-identification method of Leptospira species, a flaB gene of Leptospira was investigated and the following results were obtained. 1) HaeIII- or HindIII-restriction fragment length polymorphism (RFLP) of polymerase chain reaction (PCR) products (793 bp) of flaB gene was effectual for the classification of species of Leptospira. 2) Twenty cells of Leptospira in 1 ml of coagulated blood and 100 cells of Leptospira in 1 ml of anti-coagulated blood could be detected by flaB-PCR. These results suggested that PCR-RFLP based on the flaB gene was an efficient tool for rapid detection and identification of species of infected Leptospira from clinical specimens.

DNA Primers↗

Borrelia burgdorferi infection in UK horses.

Antibody levels (IgG and IgM) to Borrelia burgdorferi were measured in the sera and synovial fluids of UK horses. Western blotting against B. burgdorferi was also used on samples from seropositive horses. A low incidence of seropositivity was shown in horses from most parts of the UK. This increased in areas that have a high incidence of human and canine borreliosis (Norfolk and south coast). Leptospira infections of horses did not cause cross reactions in the B. burgdorferi ELISA. Most horses did not display clinical signs of Lyme disease. As with dogs and man, it is apparent that B. burgdorferi infection occurs in horses in the UK but clinical Lyme disease is uncommon.

Animals↗