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Infections in an Alpine environment: antibodies to hantaviruses, leptospira, rickettsiae, and Borrelia burgdorferi in defined Italian populations.

From 1987 to 1991, a seroepidemiologic survey for antibodies to hantaviruses, leptospira, rickettsiae, and Borrelia was conducted in selected Italian population groups. In the mountainous areas of northeastern Italy, the prevalence of antibody to hantaviruses, as detected by indirect immunofluorescent antibody (IFA) assay, was 7.1%, 4.8%, 4.3%, and 4% in 265 forestry workers, 82 rangers, 395 farmers, and 75 hunters, respectively. Among 299 Alpine soldiers, the prevalence was lower (0.7%). Of those with Hantaan antibody, the reactivity pattern using Hantaan, Puumala, and Fojnica viruses suggested a prevalence of antibody to Hantaan virus, with titers reaching levels of 128. The presence of leptospiral antibodies (by microagglutination test), which included the prevalence of antibodies to Leptospira icterohaemorrhagiae, L. bratislava, and L. saxkoening serotypes, was observed in 10-12% of the farmers and forestry workers in these Alpine mountain regions. Only a few sporadic clinical cases of leptospirosis have been reported from these regions. Antibodies to Borrelia burgdorferi (by IFA) were observed in 19% of the rangers and forestry workers, with lower values in farmers (10%) and hunters (8%). These data suggest the presence of a large number of asymptomatic infections with B. burgdorferi and the leptospires in the densely wooded areas of the Alpine Italian regions. Furthermore, the recent identification of a case of Hantaan acute nephropathy in a man living in the mountainous northeastern area of Italy confirms the presence of hantavirus in the Italian Alpine zones, especially those near the Slovenian border.

Adult↗

Sensitivity in vitro of leptospira to oxamicetin.

Oxamicetin was tested against 12 strains of parasitic leptospira each representing a different serotype and serogroup, and 8 saprophytic strains in liquid media. The results showed general susceptibility of parasitic leptospira being MIC's from 0.1 to 0.5 mug/ml. The MIC's against saprophytic strains were 10 similar to 40 mug/ml.

Anti-Bacterial Agents↗

Prevalence of antibody titers to Leptospira spp. in Minnesota white-tailed deer.

Serum samples (n = 204) from 124 white-tailed deer (Odocoileus virginianus) in northeastern Minnesota (USA) were collected from 1984 through 1989 and tested for antibodies to six serovars of Leptospira interrogans (bratislava, canicola, grippotyphosa, hardjo, icterohemorrhagiae, and pomona) using a microtiter agglutination test. Eighty-eight (43%) sera were positive at greater than or equal to 1:100 for antibodies against serovars pomona and/or bratislava; none was positive for any of the other four serovars. None of the 31 sera collected in 1984-85 was positive, whereas all 54 sera collected from 1986 through 1988 had titers of greater than or equal to 1:100. During 1989, only 34 (29%) of 119 sera had titers of greater than or equal to 1:100. Based on these results, we believe there to be wide variability in exposure of Minnesota deer to Leptospira interrogans.

Agglutination Tests↗

Serosurvey for antibodies against Brucella abortus and Leptospira interrogans in pampas deer from Brazil.

A survey for antibodies against Brucella abortus, and Leptospira interrogans was conducted on 17 pampas deer (Ozotocerus bezoarticus) from Pantanal Matogrossense (State of Mato Grosso do Sul, Brazil) and on 24 pampas deer from Parque Nacional de Emas (State of Goiás, Brazil). Antibodies against B. abortus were detected by plate agglutination, rose Bengal, and complement fixation tests; antibodies against Leptospira interrogans were detected by the microscopic agglutination test. All sera were negative for B. abortus antibodies and all deer sera from Parque Nacional de Emas were negative for L. interrogans antibodies. Four (24%) of 17 sera from Pantanal Matogrossense were positive for L. interrogans serovar (n = 2) hardjo, wolffi (n = 1) and mini (n = 1). While these diseases do not appear to be of major importance to the health status of Pampas deer, it appears that deer are reservoir for leptospirosis in one of the study areas.

Agglutination Tests↗

Cross antigenicities of Leptospira interrogans serovar copenhageni Shibaura strain for preparing biological products in Japan.

Cross protective antigenicities of Leptospira were studied with 16 antisera to 13 serovars of Leptospira interrogans and a virulent strains "Shibaura (V)", which is now considered to belong to serovar copenhageni, for challenge. The antisera of the rabbits highly immunized with various serovar live antigens were examined for leptospiricidal activity; cross immunity was recognized only in the same serogroup, Icterohaemorrhagiae. Serovar icterohaemorrhagiae lacks a prt of the antigenic components of copenhageni, but the antiserum to the former showed potent leptospiricidal activity to Shibaura (V). Anti-copenhageni immune serum absorbed with the icterohaemorrhagiae antigen did not show any protective activity to Shibaura (V).

Agglutination Tests↗

Immunological and morphological analysis of sodium dodecyl sulfate extract of Leptospira.

Sodium dodecyl sulfate (SDS) extract of Leptospira interrogans serotype icterohaemorrhagiae strain Shibaura was fractionated by rate zonal centrifugation in sucrose density gradient, and the fractions were analysed immunologically and morphologically. Two main peaks of antigenic activities were obtained. The first peak showed activities of strong indirect hemagglutination (IHA), weak complement fixation (CF), and low production of agglutinin in mice, and produced 5 precipitin lines in immunodiffusion with homologous strain antiserum. It gave only partial protection to guinea pigs. No recognizable structural components were found electron microscopically in the first peak. The second peak, on the other hand, showed no IHA activity, strong CF activity, high production of agglutinin in mice, and produced no precipitin line in immunodiffusion. It gave complete protection to guinea pigs. Membrane fragments of the enveloping sheath were found electron microscopically in the second peak. The protective activity of SDS extract of leptospira was thus intimately associated with the activityes of agglutinin production and CF, as well as with the membranous structures of the enveloping sheath.

Antigen-Antibody Reactions↗

Electrophoresis of water-insoluble leptospiral proteins and the taxonomy of Leptospira.

The cellular debris obtained after lysis of Leptospira were solubilized by sodium dodecyl sulfate and studied by polyacrylamide electrophoresis, comparing the molecular weight and relative concentration of the protein bands. The percentage similarity of 30 characteristic bands from 21 Leptospira strains, belonging to the main pathogenic and some saprophytic serogroups, was calculated; A partial matching of the derived dendrogram with the other classifications was observed.

Bacterial Proteins↗

Lipopolysaccharide biosynthesis in Leptospira.

Lipopolysaccharide is the major surface antigen of Leptospira. Variation in LPS structure is the basis for the more than 200 serovars that have been identified. Despite the importance of this antigen in immunity and diagnostics, there is relatively little known about the genetics and chemistry of leptospiral LPS, as compared to some members of the Enterobacteriaceae. The nucleotide sequence of the locus encoding enzymes for the biosynthesis of the O-antigen component of leptospiral LPS (rfb locus) has been determined for three serovars namely, L. interrogans serovar Pomona, L. interrogans serovar Hardjo subtype Hardjoprajitno and L. borgpetersenii serovar Hardjo subtype Hardjobovis. In the absence of data relating to the chemical structure or genetic tools to construct isogenic mutants in Leptospira, similarity analysis has been used to provide insight into the mechanisms by which the leptospiral O-antigen is assembled by comparison with characterized systems from other bacteria. In addition, comparison of the gene layout in each of the serovars provides an indication of the genetic basis for serovar diversity.

Enterobacteriaceae↗

[Cultivation of Leptospira in a liquid culture medium with lysed rabbit blood].

A new method of the preparation of liquid culture medium for the cultivation of Leptospira has been developed on the basis of multiple freezing and thawing of defibrinated rabbit blood. A higher productivity of the new medium in comparison with the medium used in common practice has been established. The medium having a new composition is suitable for cultivation of Leptospira and the accumulation of biomass. The advantage of the medium made is in addition to high growth properties, the its economic efficiency and availability for practical laboratories.

Animals↗

Detection of pathogenic Leptospira spp. infections among mammals captured in the Peruvian Amazon basin region.

To identify potential zoonotic reservoirs of pathogenic leptospires in the Peruvian Amazon basin, wild mammals were trapped from July 1997 to December 1998 near the city of Iquitos. After extraction of nucleic acids from animal kidneys, DNA of pathogenic leptospires was identified by polymerase chain reaction (PCR) assays using one of two primer sets, one amplifying a region of the 23S rRNA gene, and the other amplifying a gene fragment specific for Leptospira spp (G1/G2 primers). Overall, 29% (40 of 136) of the mammals tested showed evidence of renal infection by Leptospira spp., including 20% (13 of 64) of the rodents, 39% (20 of 51) of the marsupials, and 35% (7 of 20) of the chiropterans (bats). Marsupials and chiropterans were implicated as more significant reservoir hosts of leptospires pathogenic to humans than previously recognized.

Animals↗

Leptospira strains kept at the National Centre for Leptospirosis in Rome, Italy.

Since the National Centre for Leptospirosis (Department of Bacteriology and Medical Mycology, Istituto Superiore di Sanità, Rome) was established in 1956 by B. Babudieri, efforts have been devoted to identifying new Leptospira isolates and maintaining a collection of strains that today comprises 670 strains, 550 of which have been totally or partially classified, and 120 are still under study. This collection includes 23 serogroups and 156 serovars of pathogenic leptospires, and 32 serogroups and 54 serovars of saprophytic leptospires. The conventional serogroup and serovar identification, mainly based on antigenic relatedness, is tedious and time-consuming, requiring the maintenance of a comprehensive collection of serovar reference strains and the preparation of the corresponding rabbit antisera. Although considerable difficulties are encountered in the classification of leptospires at the serogroup and serovar level, this classification system is essential to obtain information on the epidemiology of leptospirosis in the different geographical areas. Serovar identification has become faster with the introduction of pulsed-field gel electrophoresis (PFGE) of large DNA fragments obtained after digestion of leptospiral DNAs with rare-cutting restriction enzymes. This technique has been successfully utilized to discriminate between closely related serovars of the Leptospira interrogans complex. We have recently used PFGE to characterize several Italian leptospiral isolates, confirming that PFGE analysis combined with microscopic agglutination test (MAT) with monoclonal and polyclonal antibodies can be used as an accurate and reliable method to compare and classify leptospires.

Animals↗

[Polymorphism of water-soluble and triton-X 100-extractable esterases and proteins from Leptospira].

Electrophoresis in gel from polyacrylamide was used to study the water-soluble intracellular esterases, triton-X 100-extracted and proteins of three saprophytic and three pathogenic strains of leptospirae belonging to different serological types. The results of investigation of a relative mobility of 27 fractions with an esterasic activity and the molecular weight of eight of them showed that their polymorphism was mainly consecuent of structural differences. In examination of proteins extracted from the cell residues with triton-X 100 there was also revealed an intertype polymorphism: by acrylamide electrophoresis with sodium dodecylsulphate they were divided into 3--10 fractions with a molecular weight of from 20000 to 200000. The results of these studies are discussed as a manifestation of biochemical individuality of leptospirae of individual serological types.

Bacterial Proteins↗

[Effect of some antibiotics and chemotherapeutic preparations on Leptospira in vitro].

It was shown in vitro with the method of successive serial dilutions that polymyxin M, thilan, streptomycin, neomycin, sodium levomycetin succinate, brilliant green, tripaphlavin, azidin, aminoacrichin, and novarsenol had the most pronounced bactericidal effect on Leptospira. Combination of the antibiotics or the chemotherapeutics apart from azidin and tripaphlavin did not increase the bactericidal effect as compared to their use alone. The clinical and collection strains of Leptospira had the same sensitivity to the substances tested. The above drugs should be tested on animals with leptospirosis.

Anti-Bacterial Agents↗

Guillain-Barre syndrome in a pediatric patient following infection due to Leptospira.

Leptospirosis is a disease with protean manifestations. We report a case of Guillain-Barre syndrome (GBS) in a pediatric patient following infection with Leptospira. Infecting Leptospira presumably belonged to serovar Copenhageni. The patient recovered completely. The possibility of GBS developing as a result of antecedent leptospiral infection should be kept in mind.

Agglutination Tests↗

[Use of culture media with sheep serum for maintenance and isolation of Leptospira strains].

A study was carried out on the effects of a culture medium prepared with sheep serum inactivated at 68 degrees C on Leptospira cultures. Good results were obtained showing that the medium with sheep serum can be used for the cultivation of Leptospira in view of the preparation of antigens for microscopic agglutination and complement fixation. In experiments on the isolation of germs from organocultures and haemocultures the proportion of positive results obtained with sheep serum was smaller than with the media containing rabbit serum.

Animals↗

Renal dysfunction associated with infection of Leptospira interrogans in a horse.

Renal failure associated with infection of Leptospira interrogans was detected in a horse. Fever, leukocytosis, pyuria, isosthenuria, and azotemia were suggestive of an inflammatory urinary tract disease. Despite persistent pyuria, no bacteria were found during routine microscopic examinations or bacteriologic culturing of urine. A fluorescent antibody examination of the urine was positive for L interrogans. Serologic testing during a 6-month period, supported an acute infection with L interrogans serovar pomona. Treatment with intravenously administered fluids and antimicrobials resulted in clinical recovery. Leptospira interrogans serovar pomona has been reported as causing fever, uveitis, or abortion in horses.

Acute Kidney Injury↗

[Anti-Leptospira antibodies in slaughterhouse workers in Ankara].

Leptospirosis is one of the zoonotic infections which is widely seen especially in tropical regions, and the primary transmission is by direct contact of water, contaminated by the secretions of animals. The aim of this study was to investigate the seroprevalence of leptospirosis in the slaughterhouse workers, who have very close contact with the animals, as a risk factor. The serum samples collected from 102 personnel (3 female, 99 male, age range: 19-66 years) who work in the different slaughterhouses in Ankara province, have been screened by means of anti-leptospira antibodies with a reference method, microscopic agglutination test (MAT). In this method, a total of 40 strains were used as antigens, including L. interrogans serotype gripptyphosa Moskva V, L. icterohaemorrhagiae Wijnberg, L. sejroe Hardjoprajitno, L. pomona Pomona, L. canicola Canicola, L. australis Jez-Bratislava and L. biflexa serotype semeranga patoc I. The evaluation was made by dark field microscopy, and the presence of agglutination in > or = 1/100 titers, were accepted as positive. As a result, in only 2 (1.96%) of the sera, anti-leptospira antibodies against L. australis Jez-Bratislava antigen were found positive, and it was detected that, these subjects were the personnel who worked in the same slaughterhouse for a mean period of 15 years. In conclusion, as 100 workers were found seronegative, the seroprevalence of leptospirosis is very low in our province.

Abattoirs↗