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PubMed1998

Leptospira interrogans antibodies in feral pigs from New South Wales.

The sera of 195 hunter-killed feral pigs (Sus scrofa), collected in New South Wales (Australia) from April to November 1995, were screened against a reference panel of 14 Leptospira interrogans serovars using a microscopic agglutination test (MAT). The panel represented those serovars previously isolated from wild and domestic mammals in mainland Australia. Antileptospiral agglutinins were detected in 20% of the sera tested and included nine L. interrogans serovars. The majority of serological reactors (63%) were to L. interrogans serovar pomona. Sera from 26% of immunoreactors cross reacted with antigens from one or more serovars. No differences were noted in the prevalence of L. interrogans antibodies between the sexes, or between pigs from areas of low and high rainfall. The implications of leptospirosis in feral pigs on the transmission of leptospires to wildlife, livestock, and humans are discussed.

Agglutination Tests↗
PubMed2003

Evidence of Leptospira interrogans infection in California sea lion pups from the Gulf of California.

Forty-two urine and 96 blood and serum samples were obtained from California sea lion (Zalophus californianus) pups from the Gulf of California during the 2000 reproductive season. Antibody prevalence to 13 serovars of Leptospira interrogans was determined by microagglutination tests (MAT); presence of pathogenic leptospires was detected by polymerase chain reaction (PCR). Samples with antibody titers > or = 1:25 or 115 bp fragments on ethidium bromidestained 1.5% agarose gels were considered positive. Antibody prevalence was 54% overall with highest prevalence against serovar cynopteri (50% of all positive reactions). Highest antibody titers (1:50) were detected against serovars cynopteri and pomona. Polymerase chain reaction products were observed in two of 42 urine samples, six of 96 blood samples, and one of 96 serum samples. Presence of PCR products in blood and serum was demonstrated in pups that were seronegative. Kruskall-Wallis tests and corresponding post hoc Tukey tests (alpha = 0.05) showed that prevalence of leptospirosis was significantly different among all rookeries. The high seroprevalence (54%), low antibody titers (maximum 1:50), absence of pups showing clinical signs indicative of the disease, and lack of recent reports of increased mortality of sea lions in the Gulf of California are suggestive of the presence of enzootic host-adapted serovars. Crowding in rookeries as well as the presence of bats and rodents on some of the islands may explain infection by L. interrogans (sensu lato) and some of the differences in seroprevalence among reproductive rookeries.

Agglutination Tests↗
PubMed1970

The susceptibility of five species of wild animals to experimental infection with Leptospira grippotyphosa.

Five species of wild animals including opossums, Didelphis marsupialis, striped skunks, Mephitis mephitis, red, Vulpes vulpes, and gray foxes, Urocyon cinereoargenteus and raccoons, Procyon lotor, were inoculated intraperitoneally with varying numbers of Leptospira grippotyphosa, organisms. Clinical signs were not detected, however, leptospiremia, leptospiruria and antibodies for the homologous organism, were demonstrated. Lesions attributed to leptospirosis were observed in liver and kidney tissue of infected animals. Infections were demonstrated in 5 of 9 opossums, 3 of 9 striped skunks, 3 of 9 red foxes, none of the gray foxes, and 9 of 9 raccoons. Therefore, it would appear that under conditions of this experiment that raccoons were most, opossums moderately, and skunks and red foxes least susceptible; gray foxes were not susceptible.

Animals↗
PubMed1974

Intracutaneous infection with Leptospira icterohaemorrhagiae (Shibaura strain) of the guinea pig.

Experimental leptospirosis with Leptospira icterohaemorrhagiae Shibaura strain was studied in guinea pigs. When the pathogen was inoculated intracutaneously to the back of the animals, localized haemorrhage was observed at the inoculated site before the appearance of general haemorrhage. The severity of the local lesion increased progressively until the 7th day of inoculation. The minimum infective dose (MID) or the 50% infective dose (ID50) of the leptospiral suspension was determined by the appearance of the macroscopic local haemorrhage 7 days after inoculation. The MID thus determined was almost comparable with the value determined by the development of general symptoms and signs by conventional ip inoculation. The number of the pathogen per ID50 varied between 6 and 35 in five experiments. The local haemorrhage was effectively protected by active or passive immunization. Microscopically, haemorrhage at the inoculated site was found mainly in the dermis, directly beneath the epidermis in particular, and accompanied with leakage of the pathogen. The pathogen was also detected abunduntly in the thickened epidermal layer covering the inoculated area as well as in the epithelial matrix of hair-follicle, probably due to the proliferation of the pathogen at the site.

Animals↗
PubMed1991

Lipids and fatty acids of Leptospira interrogans serovar copenhageni virulent strain Shibaura.

Lipids and fatty acids of Leptospira interrogans serovar copenhageni virulent strain Shibaura were analyzed by thin-layer chromatography, gas-liquid chromatography, gas-mass spectrometry and infrared absorption spectrometry. The virulent cells possessed a characteristic lipid pattern consisting of free fatty acid (FFA) (41.8%), one major unidentified phospholipid (14.8%), phosphatidylethanolamine (PE) (12.9%), cholesteryl ester (CE) (9.3%), lysophosphatidylethanolamine (LPE) (4.9%) and diphosphatidyl-glycerol (DPG) (1.1%). Various fatty acids such as hexadecanoic (26.9%), hexadecenoic (15.4%), octadecenoic (26.5%) and octadecadienoic (27.4%) acids were detected in the FFA. The fatty acid composition of the major unidentified phospholipid distinctly differed from those of other lipids including PE, LPE, DPG and CE, and comprised mainly tetradecadienoic (53.6%), tetradecatrienoic (14.0%) and octadecanoic (13.8%) acids. This phospholipid with a large amount of polyunsaturated fatty acids with chain lengths of 14 carbon atoms was detected only in the lipids of the virulent cells.

Acetone↗
PubMed1976

Serological characteristics of the Leptospira serogroup Pomona. I. Factor analysis of the reference strains.

The factor analysis of the Leptospirae of the Pomona serogroup discloses the presence of 4 main factors in this group on the basis of which 4 serovars, namely pomona, mozdok, tropica and proechimys may be accepted in this group. It is proposed that the Monjakov and the LT 1026 strains be referred to the pomona serovar. One of the established main antigens (P-4) is of the complex nature containing both thermostable and thermolabile compounds.

Absorption↗
PubMed1976

Studies on the role of sodium in the metabolism of halophilic leptospirae.

Data from studies of the metabolic mechanism of the sodium dependence of halophilic leptospirae revealed that the presence of sodium - 1) has no effect on the oxygen uptake when the microorganism is oxidizing endogeneous and exogeneous substrates; 2) is not involved in the multiplication phase; but 3) is of importance for keeping the cell structure intact.

Leptospira↗
PubMed1999

Infertility and abortion among first-lactation dairy cows seropositive or seronegative for Leptospira interrogans serovar hardjo.

OBJECTIVE: To estimate the extent to which exposure to Leptospira hardjo before or at the time of first parturition was associated with infertility and abortion during the first lactation among dairy cows that had not been vaccinated for > or = 12 months. ANIMALS: 207 first-lactation cows from a herd of 2,000 lactating cows. PROCEDURE: Cows were tested for antibodies to L hardjo within 40 days after calving. Time from calving to first breeding, time from calving to conception, number of breedings per conception, and risk of abortion were compared between cows seropositive for L hardjo and cows that were seronegative. RESULTS: For the 9 (4.3%) cows that were seropositive for L hardjo, median time from calving to conception (132.6 days) was significantly longer than time for seronegative cows (95.4 days). Cows that were seropositive were twice as likely (relative risk, 2.07) to fail to conceive as seronegative cows. Mean number of breedings required per conception for seropositive cows (3.4) was significantly higher than that for seronegative cows (2.1). The proportion of seropositive cows that aborted was not significantly different from the proportion of seronegative cows that aborted. CONCLUSIONS AND CLINICAL RELEVANCE: Exposure of nonvaccinated dairy cows to L hardjo can be associated with a subsequent reduction in fertility, as indicated by a greater time from calving to conception and higher number of breedings required per conception. The efficacy of leptospiral vaccines should be assessed to determine whether vaccination will minimize herd infertility associated with L hardjo infection.

Abortion, Veterinary↗
PubMed1997

[Characterization of immunoblots of hydrophobic outer membrane proteins Leptospira interrogans serovar lai strain 017].

Outer membrane proteins (OMP) of Leptospira interrogans serovar lai strain 017 were extracted by using Triton X-114 (TX-114). The OMP were solubilized and phase partitioned into both the hydrophilic, aqueous phase and hydrophobic, detergent phase. TX-114 did not solubilize the protoplasmic cylinder in intact organisms. The protoplasmic cylinders contained a lot of protein bands and most of the TX-114 solubilized proteins partitioned into the aqueous phase, whereas only 14 protein bands entered the detergent phase with SDS-PAGE. Detergent phase proteins were of 5 major protein bands such as 66 kd, 39 kd, 35 kd, 27 kd, and 16 kd. Immunoblotting of the material extracted with TX-114 showed that detergent phase proteins of alone 39 kd was apparently immunoblotting with antiserum against such as whole cell of 017 strain the outer envelope of 017 strain and the immunoprotective anti-017 Mb E4B7G5. The results showed that we could separate and purify the 39 kd protein to analyse the amino acid sequence for the cloning, expression and development of genetic engineering vaccines.

Bacterial Outer Membrane Proteins↗
PubMed1998

[The effect of a vaccine made from 39kd hydrophobic outer membrane protein of Leptospira interrogans on neurohumoral and red cell immunity function of the guinea pigs].

A randomized control trial was conducted to determine the immunoprotective efficacy of OmpL39. 36 guinea pigs were divided into OmpL39 group, whole leptospiral cell vaccine (WLCV) group, other proteins of Leptospira group, and negative control group (normal saline, NS). The results showed that all the guinea pigs of infected OmpL39 and WLCV still survived, but all the control guinea pigs died. Immunoprotective efficacy was 100% for OmpL39 and WLCV. OmpL39 levels were similar to WLCV levels and higher than controls (P < 0.05). These suggested that OmpL39 could be used as important immunoprotective antigen to develop the genetic vaccine of the targeting delivery system. Also it was observed that OmpL39 could produce 100% immunoprotective efficacy, have higher MAT level (> 1:3200) and regulate 5-HT, 5-HIAA, DA, NE. The 5-HT, DA, NE concentration was lower and 5-HIAA was higher than that of controls after stimulating the guinea pigs with OmpL39 (P < 0.05). The results suggested that OmpL39 genetic vaccine could produce immunity function and have an active effect on absorbing inflammation and protecting organs and tissues. Besides, the red cell immunity efficacy of the guinea pigs was changed during immunity response and the RBC-C3b RR and the RBC-ICR were increased. The RFER was increased; the RFIR was decreased. RFER/RFIR was remarkably higher than that of control (P < 0.05). These suggested that OmpL39 could increase red cell immunity function.

Animals↗
PubMed1975

Immunity to leptospirosis: Leptospira interrogans serotype pomona bacterins in cattle.

Immunity to renal leptospirosis was demonstrated in cattle vaccinated with Leptospira interrogans serotype pomona. Vaccinated cattle, challenge inoculated with virulent L interrogans serotype pomona organisms 12 months after vaccination, were resistant to renal infection; similarly inoculated, but nonvaccinated, cattle were susceptible to renal infection. Relationship was demonstrated between immunity induced in vaccinated cattle and that induced in hamsters vaccinated with a dilution of the same bacterin and later inoculated with virulent L interrogans serotype pomona organisms.

Agglutination Tests↗
PubMed2000

Lai-like leptospira from the Andaman Islands.

BACKGROUND & OBJECTIVES: Leptospirosis has been an important public health problem in the Andaman Islands since 1988. As information about the exact etiological agent is not available, the present study was undertaken to isolate and identify Leptospira from human patients. METHODS: An isolate coded AF61 was recovered from the blood of a patient clinically suspected to have leptospirosis, with fever, headache and body aches as the main symptoms. The isolation was done using Ellinghausen-McCullough-Johnson-Harris (EMJH) medium following standard procedure. The isolate was identified using microscopic agglutination test (MAT) with 'groupsera', cross agglutination absorption test (CAAT) and monoclonal antibodies. RESULTS: Agglutination tests with rabbit antisera revealed that the isolate belonged to the serogroup icterohaemorrhagiae. The CAAT results showed that it was closely related to the serovar lai. Analysis of AF61 with monoclonal antibodies confirms our observation with CAAT that it is closely related to the reference strain Lai serovar lai. INTERPRETATION & CONCLUSIONS: Serovar lai, has been associated with pulmonary haemorrhage in China and Korea. However, the strain AF61 was not isolated from a patient with pulmonary symptoms. Further studies are needed to understand the possible relationship between serovars and clinical patterns and the distribution of serovar lai and lai-like strains in Asia.

Australia↗
PubMed2000

Introduction of a rapid dipstick assay for the detection of Leptospira-specific immunoglobulin m antibodies in the laboratory diagnosis of leptospirosis in a hospital in Makassar, Indonesia.

An easy, rapid and robust dipstick assay for detection of leptospira-specific immunoglobulin M (IgM) antibodies was evaluated on 403 patients admitted for hospitalization because of fever. The clinical symptoms and signs of 35 patients were consistent with leptospirosis. The final diagnosis for the remaining patients was as follows: 136 with typhoid fever, 82 with hepatitis, 74 with malaria, 48 with infections of the respiratory tract, and 20 with fever of unknown origin. The clinical diagnosis of leptospirosis was confirmed for 24 (68.6%) patients by the combined results of the microscopic agglutination test (MAT), the reference test for leptospirosis, and of IgM ELISA, a standard laboratory test for the serodiagnosis of leptospirosis. In addition, serum specimens from 8 (2.2%) patients with a final clinical diagnosis other than leptospirosis were found to be positive in MAT and/or IgM ELISA. Compared with the results of MAT and IgM ELISA a sensitivity of 91.6% and specificity of 93.6% was calculated for the dipstick assay. Most of the serum samples from the laboratory confirmed patients gave a moderate to strong staining intensity of the antigen band of the dipstick and were easy to read. The results demonstrate that the dipstick assay is convenient to use and allows the rapid and accurate confirmation of patients with clinical suspicion of leptospirosis in areas where the disease is endemic.

Antibodies, Bacterial↗
PubMed2001

Sensitivity of Borrelia and Leptospira to Quinupristin-dalfopristin (Synercid) in vitro.

In vitro activity of Quinupristin-dalfopristin (Synercid) against seventeen isolates of Borrelia burgdorferi and two representatives of Leptospira spp. was investigated. MICs ranged from 0.03 to 0.125 for B. burgdorferi and 0.125-0.25 microg/ml for Leptospires. Time killing studies carried out with 2 MIC demonstrated U 3 log(10)-unit killing after 72 h, showing a significant activity against spirochetes, though at a lower level than other antibiotics in use in the therapy of Lyme disease and leptospirosis.

Anti-Bacterial Agents↗
PubMedDate not supplied

[Leptospira interrogans serovar Pomona: detection of antigenic differences among 3 regional isolates from cattle and a reference strain].

Leptospirosis is a zoonosis caused by Leptospira interrogans. This disease is diagnosed by quantification of specific immunoglobulins in serum by the microagglutination test (MAT). The aims of this research were: a) to compare the protein profiles of 3 clinical isolates of bovine leptospirosis with the reference strain used for the MAT, and b) to identify the immunodomain antigens of the regional isolates through PAGE and immunoblotting techniques of bovine sera from infected, vaccinated and MAT-negative animals. Coomassie-blue stained gels revealed extensive protein similarities between pathogenic and reference strain. Most infected (8/10) and vaccinated animal sera (4/7) showed by immunoblotting a similar reactivity against the proteins from pathogenic leptospires, with a strong band of 25-30 kDa which was not detected in the reference strain. The lack of correlation between MAT and immunoblotting techniques for infected animals could be due either to the infection stage at which the diagnosis was made or to the immunoglobulin isotype involved in the response. Results obtained would confirm the antigenic differences between the 3 isolates and the reference strain.

Agglutination Tests↗
PubMedDate not supplied

[Detection of anti-Leptospira antibodies in sera of patients in the latex agglutination test].

The results of the preliminary evaluation of the sensitivity and specificity of the newly developed diagnostic test based on the determination of genus-specific antibodies to leptospires in the latex agglutination test, are presented. This test makes it possible to detect anti-Leptospira antibodies of any serogroup. The advantages of the developed test have been determined.

Antibodies, Bacterial↗
PubMed1975

[The result-sequence method (sequence analysis) in leptospira research. 3. Communication: The bilateral sequential test (de Boer, Armitage) for testing the difference of mean values of two binomial distributions (author's transl)].

The influence of the liver-activated and non-activated cytostatic drug Cyclophosphamid on the respiration of Leptospira biflexa semaranga Veldrat S 173 was tested in three different concentrations by group-sequential testing of two relative frequencies in a two-sided test-reading. The conditioned probability theta = 0.82 results from thetan = 0.40 and thetaa = 0.75. On account of a practicable sample size, we choose theta' = 0.80 (activated form superior) and theta'' = 0.20 (non-activated form superior). The test-adjusted level of significance is alpha = 0.05 less than beta = 0.10. The expected values for the number of discordant pairs are Etheta' = Etheta'' = 13, Etheta = 1/1 = 14 and Ethetamax = 16. The acceptance inspection performed by control chart and tabulated reference figures results - in comparison with conventional procedures - in savings of discordant pairs of 65 per cent (concentration 10(-4) g/ml) in a decision in favour of activated form, of 65 per cent (concentration 10(-8) g/ml) in an equal efficiency of both states of Cyclophosphamid and of 10 per cent (concentration 10(-11) g/ml) in a non-significant difference. Taking into consideration all unrestrictedly selected pairs, the duration of the experiment takes only 4.25 months instead of 6.75 if the statistical data analysis is not performed conventionally, but group-sequentially instead.

Cyclophosphamide↗