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Evaluation of an indirect hemagglutination test for the diagnosis of human leptospirosis.

A presumptive hemagglutination test for the serological diagnosis of leptospirosis in humans is described. The antigen was prepared from a soluble alcohol extract of an andamana strain sorbed to human O-negative erythrocytes and preserved by pyruvic aldehyde fixation. In this study, the overall sensitivity of the hemagglutination test was 92% in contrast to 69% for the presumptive slide agglutination test. The specificity was 95% for the hemagglutination test in comparison with 83% for the slide test.

Agglutination Tests↗

Impact of proficiency testing on results of the microscopic agglutination test for diagnosis of leptospirosis.

A proficiency testing scheme for the leptospirosis microscopic agglutination test was provided to 37 laboratories in 23 countries in 2002 (round 1) and to 60 laboratories in 34 countries in 2003 (round 2). Thirty-four laboratories participated in both rounds. Each panel consisted of five rabbit serum samples, four of which were antisera raised against pathogenic serovars of Leptospira. One of these samples was a mixture of two different antisera. The rates of false-negative results, calculated on the basis of the assumption that serovars within a serogroup will cross-react, were 11% for round 1 and 14% for round 2. There were regional differences in the rates of false-negative results. The titers reported by laboratories testing for the same sample with the same serovar varied widely. Laboratories that had previously participated in round 1 reported fewer false-negative results in round 2 than new participants (10 and 21%, respectively [P = 0.002]) and reported 0.56 false-negative results per participant, whereas new participants reported 1.23 false-negative results per participant (P = 0.041). Laboratories that had previously participated also reported fewer false-negative results in round 2 than in round 1 when samples common to both rounds were tested (5 and 15%, respectively [P = 0.028]). The titers reported by the new participants were, on average, lower than those reported by the laboratories that had participated previously (P = 0.019) and were significantly more variable (P = 0.001). Analysis of these results suggests a positive impact of proficiency testing on the testing performance of the participating laboratories.

Agglutination Tests↗

Benign leptospirosis: first reported outbreak in British Isles due to strains belonging to the Hebdomadis serogroup of Leptospira interrogans.

The clinical and epidemiological features associated with the first reported outbreak in the British Isles of benign anicteric leptospirosis, due to strains belonging to the hebdomadis subgroup, are described. Four cowherdsmen working in two dairy farms in Surrey developed a febrile illness which was not associated with jaundice, but aseptic meningitis was a feature. Microscopic agglutination tests with formolized antigen suspensions revealed a significant rise in the agglutinin titres against various serotypes belonging to the hebdomadis serogroup of Leptospira interrogans. The probable source was eventually traced to the cattle, which showed serological evidence of infection with the same serogroup.

Adult↗

Failure of penicillin prophylaxis in laboratory acquired leptospirosis.

A laboratory technician developed leptospirosis following accidental inoculation, despite prompt administration of parenteral penicillin by an accepted regimen of post-exposure prophylaxis. Another technician was similarly exposed and was given doxycycline; no illness or serological conversion followed. The implications of these cases are discussed and recommendations made for post-exposure chemoprophylaxis with doxycycline.

Accidents, Occupational↗

Decision support models of leptospirosis in dairy herds.

Following the results of a survey which found that 61 per cent of dairy farmers felt that they needed more information about leptospirosis, and the strategies for its control and the costs and benefits involved, this paper describes the construction and preliminary results of two models of the disease intended to help explore the risks and financial implications of Leptospira interrogans serovar hardjo infection for dairy producers.

Animals↗

The use of methanol extract of Leptospira interrogans in complement fixation tests for leptospirosis.

Methanol extracts were obtained from L. interrogans serovars icterohaemorrhagiae and canicola and L. biflexa serovar patoc. Human sera from 167 normal individuals and 40 patients with different infectious diseases tested by complement fixation tests showed negative reactions. Sera from 100 patients with a suspicion of leptospirosis were tested by complement fixation tests and microscopic agglutination reactions. Agreement of 84% was found for those two reactions. Positive microscopic agglutination tests at a dilution 1:20-1:400 with negative complement fixation tests were observed in 5% of patients and negative microscopic agglutination with complement fixation tests in the range of 1:20-1:1280 were observed in 11% of the cases.

Agglutination Tests↗

Potential application of low-stringency single specific primer-PCR in the identification of Leptospira in the serum of patients with suspected leptospirosis.

In this study we tested the potential use of low-stringency single specific primer-PCR (LSSP-PCR) for genetically typing Leptospira directly from biological samples. Serum samples obtained from 29 patients with clinically suspected leptospirosis were amplified by specific PCR, using the previously selected G1 and G2 primers. The PCR products of approximately 300 bp were subsequently used as a template for LSSP-PCR analysis. We were able to produce genetic signatures from the leptospires present in the human samples, which permitted us to make a preliminary identification of the infective serovar by comparing the LSSP-PCR profiles obtained directly from serum samples with those from reference leptospires. Thus, LSSP-PCR has the potential to become a useful diagnostic tool for identifying leptospires in biological samples without the need for bacteria isolation and culture.

Bacterial Typing Techniques↗

Acute renal failure of leptospirosis: nonoliguric and hypokalemic forms.

Acute renal failure induced by leptospirosis was studied in 56 patients. A higher frequency of nonoliguric renal failure was observed with lower morbidity and mortality rates than in oliguric forms. In addition, 45% of the patients in this series were hypokalemic, and no hyperkalemic patients were seen. A prospective study in 11 patients showed an initially elevated urinary fractional potassium excretion that fell simultaneously with the high urinary fractional sodium excretion and the urinary K/Na ratio, suggesting an increased distal potassium secretion due to an increased distal sodium delivery consequent to functional impairment of the proximal reabsorption of sodium.

Acute Kidney Injury↗

Unusual electrocardiographic abnormality in leptospirosis: case reports.

Unusual ECG changes of marked ST segment elevation in leads V1 to V3 are reported in four cases of severe icteric leptospirosis for the first time. These changes normalized rapidly with initiation of therapy and recovery in three patients. One patient died within hours of admission. The causes for the changes are not clear.

Adult↗

Leptospirosis in equine fetuses, stillborn foals, and placentas.

Leptospirosis was diagnosed in 51 equine fetuses and 16 stillborn foals with gestational ages from 3 1/2 to 11 months. Diagnosis was based on one or more of the following: positive fetal antibody titer, positive fluorescent antibody test, demonstration of spirochetes in kidney and/or placental sections stained by the Warthin-Starry technique, high leptospiral titers in aborting mares, or isolation of Leptospira spp. from fetal organs. Gross lesions were observed in 80.3% of the fetuses, stillborn foals, and placentas. Gross placental lesions included nodular cystic allantoic masses, edema, areas of necrosis of the chorion, and necrotic mucoid exudate coating the chorion. The liver (23 cases) was enlarged, mottled, and pale to yellow. The kidneys (seven cases) were swollen and edematous with pale white radiating streaks in cortex and medulla. Microscopic lesions were observed in 96% of fetuses, stillborn foals, and placentas. Placental lesions consisted of thrombosis, vasculitis, mixed inflammatory cell infiltration of the stroma and villi, cystic adenomatous hyperplasia of allantoic epithelium, and villous necrosis and calcification. Fetal lesions included hepatocellular dissociation, mixed leukocytic infiltration of the portal triads, giant cell hepatopathy, suppurative and nonsuppurative nephritis, pulmonary hemorrhages, pneumonia, and myocarditis. Spirochetes were demonstrated with the Warthin-Starry stain in the allantochorion and/or kidney of 69 of the 71 cases. Using the direct fluorescent antibody technique, 56/60 cases tested positively for leptospires. Leptospires were isolated from fetal tissues in 20/42 cases. Sixteen of the isolates were identified by restriction enzyme analysis as Leptospira interrogans serogroup Pomona serovar kennewicki; case Nos. 36 and 41 were serovar grippotyphosa.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Leptospirosis complicated by severe aortic stenosis.

A previously well 62-year-old male from North Queensland presented with leptospirosis featuring fever, renal failure, hepatitis and pulmonary haemorrhage. Management was greatly complicated by severe and previously unrecognized aortic stenosis with a peak valve gradient of 125 mmHg. A successful outcome followed careful haemodynamic management and treatment of the infective illness with subsequent valve replacement.

Aortic Valve↗

An improved immunohistochemical diagnostic technique for canine leptospirosis using antileptospiral antibodies on renal tissue.

The purpose of this study was to compare the immunoreactivity in canine renal tissues stained with antisera specific for 3 leptospiral antigens and those processed with traditional staining methods. In addition, immunoglobulin staining was done on tissues with immunoreactivity to leptospiral antigens. Formalin-fixed renal sections from 12 dogs with chronic interstitial nephritis suspected or proven to have leptospirosis (6 dogs with silver-stained leptospires and 6 dogs in which silver-stained leptospires were not detected) were used. Antibodies consisted of a monoclonal antibody to Leptospira kirschneri serovar grippotyphosa lipopolysaccharide (LPS) and 2 polyclonal antibodies to outer membrane proteins, including OmpL1, a leptospiral porin, and LipL41, an outer membrane lipoprotein. The murine monoclonal antisera against LPS (F71C2-1) had the most abundant and consistent immunoreactivity. Immunoreactive areas were present in 6 of 6 sections positive by silver staining and included extracellular granular debris in intertubular areas, debris in macrophages, organisms in tubular lumina, and cytoplasmic granules in tubular epithelia. Antisera with specificity for the outer membrane proteins OmpL1 and LipL41 detected only intact organisms in tubular lumina. Immunoreactivity to OmpL1 (polyclonal 338) occurred in 4 of 5 sections positive by silver staining, but immunoreactivity to LipL41 (polyclonal 813) occurred in only 1 of 6 silver-positive sections. Each of the kidney sections in which leptospiral antigens were detected by immunohistochemistry also was positive by silver staining. Sections negative by silver staining were also negative by immunostaining. Although immunohistochemistry did not enhance sensitivity, amplification of signal by secondary antibody and hematoxylin counterstaining improved the ease of diagnosis and allowed better evaluation of tissue morphology than did silver staining methods. IgG was the most abundant immunoglobulin. IgG immunoreactivity occurred predominantly in plasma cells within interstitial infiltrates. Interstitial infiltrates contained abundant immunoreactivity to LPS, but immunoreactivity to OmpL1 and LipL41 was not noted.

Animals↗

Serological survey of leptospirosis in sows with premature birth and stillbirth in Chiba and Gunma prefectures of Japan.

In 1996 and 1997, the seroprevalence against Leptospira in parturient sows with premature birth or stillbirth from two herds was investigated. In three out of four sow serum samples obtained in Gunma Prefecture, the antibody titers to Leptospira interrogans serovar copenhageni (M20) were higher than 10,000 (the reciprocal of the serum dilution). Furthermore, the antibody titers to L. interrogans serovar canicola (Hond Utrecht IV) were significantly high in the three sows and the titers ranged from 1,000 to 3,000. In sows obtained in Chiba Prefecture, significantly high antibody titers to serovar copenhageni (M20) were confirmed in eight out of 40 sows, and antibody titers greater than 10,000 in six of them. Significantly high antibody titers to L. interrogans serovar icterohaemorrhagiae (RGA) and L. canicola (Hond Utrecht IV) were confirmed in four and 18 out of the 40 sows, respectively, compared with the titers to the other serovars. These findings may indicate the prevalence of leptospirosis in pig herds in both Gunma and Chiba Prefectures.

Abortion, Veterinary↗

Acute leptospirosis in a triathlete.

We report the case of a 38-year-old male patient with acute leptospirosis. The most probable cause of infection was repeated and prolonged exposure to contaminated river water (Neckar and Enz rivers) while preparing for participation in long-distance triathlon (swimming, biking, and running) competitions.

Acute Disease↗

[Involvement of the nervous system in leptospirosis. I. Evaluation of neurologic aspects].

From January 1st up to September 30th 1990, 77 patients with leptospirosis confirmed by laboratory studies were admitted at the Infectious and Parasitic Diseases Service of the Hospital das Clínicas of the Universidade Federal de Pernambuco. Clinical manifestations had sudden onset with presence of fever, headache, chills and muscle pains. Plurisystemic involvement was observed both in the icteric and in the non-icteric patients. The neurological exam was abnormal in 70 of the patients (90.91%). Neurological findings were essentially variable. Though in a transitory form, they allowed the observation of the following clinical forms: meningoencephalitis and polyneuritis in association, meningoencephalitis, polyneuritis, and subarachnoid hemorrhage.

Central Nervous System Diseases↗

[Hematologic changes in leptospirosis].

30 cases of leptospirosis admitted to the Clinica de Doenças Infecciosas e Parasitárias do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo were studied for blood count alterations. 16 patients (53.3%) had a normal white blood cell count at the moment of admission. 12 patients (40%) presented a high white blood cell count and 2 (6.6%) had a low count. 29 patients (96.6%) had a high proportion of neutrophils and 25 patients (83.3%) presented a high number of immature forms. 24 patients had anemia. Thrombocytopenia was present in 26 patients (86.6%). The most characteristic changes found in bone marrow aspirate were the alterations of M:E ratio (myeloid-erythroid ratio) due to relative and/or absolute hyperplasia of the myeloid series, and/or relative and/or absolute hypoplasia of the erythroid series; erythropoiesis was predominantly micro-erythroblastic in many patients; mild to moderate plasmocytosis was found; and, regarding the interstitial series, increased macrophagic activity was noted. There was no direct correlation between the number of megakaryocytes and the blood platelet count, but there was a direct correlation between the bone marrow platelet production and blood platelet count. We believe that it is very difficult to have a good idea of the dynamic mechanisms that lead to medullar platelet production in the presence of platelet consumption, through a random test of the bone marrow.

Adolescent↗

[Outbreak of human leptospirosis by recreational activity in the municipality of São José dos Campos, São Paulo. Seroepidemiological study].

An outbreak of human leptospirosis due to recreational activities occurred at São José dos Campos, São Paulo, Brazil in November 1987. It involved a group of persons who had participated in a gathering in a suburb club which had a swimming pool fed with natural water. Epidemiological investigation was carried out and laboratory tests from the patients were done. It was observed that a high prevalence of the pomona serotype (91%) was found in the serological analyses, while the presence of the agent of the infection could not be found in the water club swimming pool.

Agglutination Tests↗