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Severe Leptospira autumnalis infection was associated with acute acalculous cholecystitis and pancreatitis in a 66-year-old man. He was successfully treated with antimicrobial agents and supportive therapy, including hemodialysis. We review these uncommon manifestations and the effectiveness of antimicrobial therapy in advanced leptospirosis.
The authors report the high-resolution computed tomography findings in five patients with diffuse pulmonary hemorrhage caused by serologically proven leptospirosis. The main findings consisted of extensive ground-glass opacities and patchy areas of airspace consolidation.
Leptospirosis is a worldwide zoonosis affecting wild and domestic mammals. It is most commonly transmitted to humans by indirect contact with infected animals. Vascular injury, mainly capillary damage, and hemorrhagic diathesis are prominent features in the affected organs. The most severe form of the disease presents with high fever, intense jaundice, hemorrhagic diathesis, hepatic and renal dysfunction, mental status changes, and cardiovascular collapse. Focal or diffuse areas containing alveoli filled with erythrocytes characterize the pulmonary involvement. These latter conditions, although unusual, are associated with significant mortality.
Isolates of Leptospira from two human cases of leptospirosis in Denmark and France were studied using DNA-DNA relatedness, G + C content, 16S rRNA gene sequence data and pulsed-field gel electrophoresis. These isolates differed from previously described species of Leptospira and are defined as Leptospira broomii sp. nov. The type strain is 5399T (= ATCC BAA-1107T = KIT 5399T).
Leptospirosis in the Philippines is an underrepresented disease. To achieve an accurate means of serodiagnosis, we demonstrated antibodies to the prevalent Leptospira serovars in sera of 71 patients from three major hospitals in Manila by the microscopic agglutination test and Western blot analysis. Sera of 53 patients contained antibody against 8 serovars poi, tarassovi, manilae, pyrogenes, australis, grippotyphosa, javanica, and autumnalis.
A 6.5-year-old male German Shepherd acutely developed renal and hepatic disease. Serology revealed high concentrations of antibodies against Leptospira copenhageni, and a presumptive diagnosis of leptospirosis was made. The dog was successfully treated with antibiotics and supportive care over a 12-day period. Sixty-two days after the initial presentation, alopecia predominantly involving the dorsum and perineal areas developed. The skin lesions expanded over a 20-day period. Histology revealed generalized calcinosis cutis with follicular atrophy. An injection of 0.01 mg kg-1 dexamethasone suppressed serum cortisol concentrations. No treatment was given and lesions resolved over the following 30 days. This is the third case of generalized calcinosis cutis that has developed in an adult dog after severe systemic disease. Both previous cases developed calcinosis cutis in association with blastomycosis. To the authors' knowledge, this is the first report of generalized calcinosis cutis in an adult dog in association with a presumptive bacterial infection.
A case of panuveitis as a result of leptospiral infection in an 8-year-old Jack Russell Terrier is reported. Mild hyphema, aqueous flare, and partial serous retinal detachments were noted in both eyes on initial ophthalmic examination. The serum microscopic agglutination testing for Leptospira serovarieties revealed titers of 1 : 12 800 for Leptospira canicola and 1 : 200 for Leptospira icterohemorrhagiae. With medical therapy, the panuveitis resolved and functional vision was regained. Four weeks after the initial presentation, convalescent titers were 1 : 200 for L. canicola and 1 : 100 for L. icterohemorrhagiae. The incidence of leptospirosis, along with difficulties diagnosing, and current medical therapies for this disease are also discussed.
AIMS: Andaman and Nicobar Islands in India has a century long history of human leptospirosis. Several isolates have been recovered over the years from different locations. The present study was undertaken to understand the clonal relationship between all these pathogenic leptospires recovered from these islands. METHODS AND RESULTS: Arbitrarily primed polymerase chain reaction (AP-PCR) was employed to genetically characterize 40 isolates recovered during 1995--2001 and their fingerprints were compared with those of 26 reference strains of known genetic and serological affinities. Sequences of PCR-amplified products from representative isolates were compared with those of different strains belonging to seven genospecies. AP-PCR fingerprints revealed that 32 of the 40 isolates were clonal in nature and fingerprints of all the isolates matched with known reference strains of pathogenic Leptospira interrogans sensu stricto. Comparison of sequence data of PCR amplified products of reference strains and isolates also corroborated these findings. CONCLUSIONS: The study revealed that 80% of the isolates recovered from these islands were clonal in nature and all the isolates taken in the study belonged to Leptospira interrogans sensu stricto. SIGNIFICANCE AND IMPACT OF THE STUDY: An extension of the study in animal population would help in understanding the transmission dynamics of this commonly circulating clone in these islands, which in turn might help in effective control of this public health problem.
Skin reactivity to tuberculin has been studied during the course of experimental leptospirosis in guinea pigs. A depression of the delayed hypersensitivity to tuberculin was demonstrated in the infected animals. The depression was most pronounced when icterus had developed. The depression was not correlated with the amount of infectious units administered or with the demonstration of live leptospirae in the peritoneal cavity. In the infected animals there was no correlation between the initial and the final skin tests which is in contrast to findings in the control group.
Case records of 36 dogs with confirmed leptospirosis diagnosed at the New York State College of Veterinary Medicine from 1980 to 1995 were reviewed retrospectively, and clinical, serological and pathological findings were recorded to characterise the epidemiology of this disease in upstate New York. Titres were directed predominantly against serovars grippotyphosa and/or pomona in 31 of 34 dogs. Convalescent titres were measured for 53 per cent of dogs. The most common clinical presentation was acute renal failure. Increased liver enzyme activity was documented in 22 of 36 dogs. It is clear from this study that Leptospira pomona and grippotyphosa are important pathogens capable of causing severe renal and hepatic injury in dogs.
Kidneys from cattle at slaughter were examined for the presence of leptospires. Of 218 (8.3%) kidneys leptospires were isolated from 18; all were identified as Leptospira interrogans serovar hardjo. None of the leptospire-infected kidneys had histopathological lesions indicative of leptospirosis and leptospires were demonstrated in only 2 by immunogold silver staining. Leptospires infected kidneys remained viable for at least 21 days when stored at 4 degrees but became non-viable within 14 days when stored frozen at -15 degrees.
A simple technique is described for drying sera on washed, dry sand or ordinary sugar cubes for the sero-diagnosis of leptospirosis. The results are shown to be very similar to those obtained with fluid sera or sera dried on filter paper discs. Sera adsorbed on sand or absorbed in sugar eliminated some of the problems associated with sera dried on paper. This method is suitable for use in the field and is expected to be of value in epizootiological studies where contamination and chemical denaturation of fluid serum samples held without refrigeration is a problem.
An indirect hemagglutination test for the diagnosis of leptospirosis is described; the test uses a soluble antigen from serotype patoc to sensitize sheep erythrocytes which are then fixed with glutaraldehyde. Evaluation of this procedure indicates that it is more reliable than the conventional macroscopic agglutination test and, in contrast with both microscopic and macroscopic agglutination tests, is positive only with sera from persons with current leptospiral illness. The test is simple and convenient and sensitized fixed cells may be stored for at least a year. In comparison with the macroscopic and microscopic tests, only a single antigen is required.
The objective of the present study was to investigate the usefulness of a recombinant flagellar protein, FlaB, of Leptospira interrogans serovar pomona in the serodiagnosis of leptospirosis by the fluorescence polarization assay (FPA). The recombinant protein FlaB was purified to homogeneity by a combination of nickel-nitriloacetic acid agarose chromatography, electrophoresis, and electroelution. Purified FlaB was labeled with fluorescein isothiocyanate (FITC). Western blotting was performed by using bovine sera with microscopic agglutination test (MAT) titers of antibodies against L. interrogans serovar pomona and L. bergpetersenii serovars hardjo and sejroe to confirm the antigenicity of FlaB. Western blot analysis demonstrated that labeled as well as unlabeled FlaB was recognized by the positive sera tested, indicating the broad serovar cross-reactivity of this protein. It also indicated that labeling with FITC did not affect the antigenicity. By using FITC-labeled FlaB as a tracer antigen, a homogeneous FPA was developed to detect antileptospiral antibodies in bovine sera. A population of 208 MAT-positive and 208 MAT-negative serum samples was tested by FPA. The FPA cutoff was determined by receiver operating characteristic analysis. By FPA, 83. 7% of the MAT-positive serum samples were positive and 81.2% of the MAT-negative serum samples were negative. Compared to the results of MAT, the positive predictive value of FPA was 81.7% and the negative predictive value of FPA was 83.3%. The FPA is a simple and rapid technique for the detection of anti-Leptospira antibodies.
An assay device for the rapid detection of Leptospira-specific immunoglobulin M (IgM) antibodies in human sera is presented. The sensitivity (85.8%) and specificity (93.6%) of the assay compared well (91.9% agreement) with those of an IgM enzyme-linked immunosorbent assay routinely used in the serodiagnosis of leptospirosis. The sensitivity of the assay varied with the stage of the disease. The assay uses stabilized components and is simply performed by the addition of serum and sample fluid to the sample well of the assay device. The assay is read after 10 min, and a positive result is obtained when staining of the test line is observed.
Whole blood and plasma from animals in the acute stage of leptospirosis contained a toxic factor which produced a cytopathic effect on fibroblastic L cell monolayers. Firm adsorption of cytotoxic factor (CTF) to L cells occurred within 1 h. The highest titer of CTF in plasma was reached at 24 h and declined after 48 h after the inoculation of leptospires. Toxic effects were also obtained with intracerebral inoculation of mice with plasma containing CTF. Mice showed signs of motor instability and muscular spasms shortly after inoculation with CTF. Death usually occurred within 1 h.
The recent availability of the complete genome sequences of Leptospira interrogans, the agent of leptospirosis, has allowed the identification of several putative virulence factors. However, to our knowledge, attempts to carry out gene transfer in pathogenic Leptospira spp. have failed so far. In this study, we show that the Himar1 mariner transposon permits random mutagenesis in the pathogen L. interrogans. We have identified genes that have been interrupted by Himar1 insertion in 35 L. interrogans mutants. This approach of transposon mutagenesis will be useful for understanding the spirochetal physiology and the pathogenic mechanisms of Leptospira, which remain largely unknown.
A passive microcapsule agglutination test for the diagnosis of leptospirosis was developed by utilizing chemically stable microcapsules instead of sheep erythrocytes. In the test, sonically disrupted antigens of leptospira were sensitized to microcapsules treated with glutaraldehyde. Compared with the microscopic agglutination test, the passive microcapsule agglutination test showed a relatively genus-specific tendency and a 4- to 32-fold-higher sensitivity. The sensitized microcapsule antigens were stable for at least 1 year. The microcapsules coupled with mixed antigens can be used as a serodiagnostic screening test for diseases caused by various types of leptospira. The test, which is very simple and reproducible and requiring no specific training, can be employed easily as a routine test in diagnostic laboratories.