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Reaction of monoclonal antibodies with species specific determinants in Leptospira interrogans outer envelope.

A set of 24 monoclonal antibodies (MABs) was produced against an outer envelope preparation from Leptospira interrogans serovar copenhageni. The MABs reacted in enzyme immunoassay with species-specific determinants of an antigen in the leptospiral outer envelope (OE) of pathogenic but not of saprophytic species of Leptospira. The MABs did not agglutinate whole leptospires, nor could they opsonise homologous leptospires for phagocytosis by mouse macrophages or protect new-born guinea-pigs against lethal infection. The MABs reacted by Western blotting with a 35 x 10(3)-mol-wt band in OE separated on SDS-polyacrylamide gels, and also reacted with other bands to a lesser extent. The determinants to which the MABs were directed were localised in the leptospiral OE by immunogold labelling techniques.

Agglutination Tests↗

The bacteriological prevalence of leptospiral infection in cattle and buffaloes in West Malaysia.

A cross-sectional bacteriological survey of cattle in West Malaysia revealed 14.4% (32/222) had leptospiral infection. Isolates were obtained from all except one herd with prevalence of infection in herds ranging from 0-44.8%. A small number of buffalo urine samples were examined and all of them were found to be negative. A leptospiral isolate obtained from a bovine kidney proved to be a new serovar of Leptospira interrogans and the name unipertama was assigned to it. Six other leptospiral serovars were isolated, namely canicola, australis, javanica, ballum, pomona and hardjo. All six serovars were isolated for the first time in cattle in Malaysia. Cattle in Malaysia appear to be the maintenance host for serovar hardjo. The presence of the other serovars in cattle was probably due to contact with the maintenance hosts, pigs for serovar pomona and rodents for the other three serovars. It appears that the epidemiology of leptospiral infection in cattle in Malaysia is similar to that reported overseas.

Animals↗

Detection of leptospiral DNA by PCR.

An EcoRI fragment (1.2 kb) which is highly conserved among Leptospira interrogans isolated in Korea was cloned into pBluescript vector from L. interrogans serovar lai WH20. The EcoRI fragment was sequenced, and a pair of primers (LP1 and LP2) was designed for PCR assay. PCR amplification of target DNA obtained from cultured L. interrogans showed that 274 bp could be detected when as little as 100 fg of leptospiral genomic DNA was used in the reaction mixture. No amplification of DNA was detected from DNA of Leptospira biflexa serovars patoc and sau paulo, Borrelia burgdorferi, Staphylococcus aureus, Escherichia coli, and Salmonella typhimurium. Amplification of 274-bp target DNA could be detected in DNA samples purified from 500 microliters of blood collected from experimentally infected gerbils 2 days after infection, while antibodies to L. interrogans could be detected by the microscopic agglutination test 7 days after infection. The specificity and high sensitivity of the test provided valuable tools for the early diagnosis of leptospirosis.

Agglutination Tests↗

Discovery of the causative organism of Weil's disease: historical view.

On January 20, 1915, Inada and Ido announced the discovery of the causative agent of Weil's disease. Subsequently, on February 13, 1915, they published the first paper on the discovery of the causative organism (a new species of Spirochaeta) of Weil's disease. Besides discovering the causative organism of the disease, Inada and colleagues clarified the pure culture in medium, and determined the source and route of the infection, its pathology and morbid anatomy; the distribution of the organism in various organs and tissues; the excretion of the spirochete, and its division, filterability, and morphological characteristics; and the clinical picture, laboratory findings, diagnosis, prophylaxis, and treatment of the disease. These studies were conducted by Inada, Ido, Kaneko, Hoki, and Ito, in the years 1914 to 1915. In the early investigation of leptospirosis, Inada and colleagues played a prominent part. We would like to remember these remarkably complete and definitive original achievements on leptospirosis made by Inada and colleagues.

Animals↗

[The serovars of Leptospira interrogans isolated from cases of human leptospirosis in São Paulo, Brazil].

Eighteen strains of L. interrogans isolated from human cases were serotyped by the agglutinin-absorption test at Instituto Adolfo Lutz in São Paulo, Brazil. Fourteen were identified as serovar copenhageni (icterohaemorrhagiae serogroup), 2 as canicola (canicola serogroup), 1 as castellonis (Ballum serogroup) and 1 as pomona serogroup (serovar not yet defined). The frequency of serovar copenhageni in 100% of the isolates in icterohaemorrhagiae serogroup is emphasized and more studies to verify the real serovars prevalence as subsidy to the epidemiology of this infection are suggested by the authors.

Agglutination Tests↗

Human leptospirosis in the Mekong delta, Viet Nam.

To estimate the seroprevalence of human leptospirosis in the Mekong delta in Viet Nam, an epidemiological survey was conducted in the province of Tien Giang, which is representative of the socioeconomic activities of the region (rice growing and cattle breeding). A cross-sectional study included 35 clusters representing 1400 people randomly selected and aged 15-60 years. Sex, age, occupation, contact with animals, type of water supply, and individual habits were recorded. Leptospiral agglutinins were detected by the microagglutination test, with a battery of 22 live antigens representing the main pathogenic serogroups of Leptospira species and additional local strains. 263 sera (18.8%) gave positive results and 41 (2.9%) had a titre of agglutinins > or = 400, suggesting recent infection. No significant difference was found between females and males. The distribution of seroprevalence was homogeneous throughout the population studied, with the exception of the 15-25 years age group, in which leptospiral antibodies were less frequent. Fifteen serogroups were found, the most prevalent being Bataviae (21.7%), Panama (15.2%), Icterohaemorrhagiae (13.7%) and Australis (8.7%). No significant link between leptospiral seropositivity and professional activities or contacts with animals was found, indicating that leptospirosis in the Mekong delta may be considered as an environmentally linked disease. Leptospirosis is known to be endemic in south-east Asia, and these data demonstrated the high level of circulation of leptospires and the potential importance of leptospiral infections among the rural population in this area.

Adolescent↗

Prevalence and clinical significance of leptospiral antibodies in pigs in England.

The results of leptospiral serology conducted on porcine sera at the Central Veterinary Laboratory, Weybridge from 1971 to 1978 were collated and analysed. Sera were available from diagnostic submissions (1622) and from pigs intended fro export (5031). A random sample of 792 sera from 34 herds was also examined. The overall prevalence of infection was low. Copenhageni titres were present in 7.9 per cent of diagnostic submissions and a statistical association with jaundice was shown in piglets and adults. The introduction of Australis serogroup antigens in 1978 revealed the serological prevalences to lora and bratislava to be in excess of 20 per cent. However the causative serotype has yet to be identified. Leptospiral infection was not a significant cause of abortion in pigs. The pattern of leptospiral infection in pigs in England is considerably different from that occurring in other countries and it is considered that free-living maintenance host populations constitute the primary sources of infection.

Abortion, Veterinary↗

Leptospiral carriage by mice and mongooses on the island of Barbados.

Leptospirosis is a zoonotic disease, maintained by chronic infection of the kidneys of reservoir animals, usually small mammals. Infection in humans is acquired from direct or indirect exposure to the urine of infected animals. Leptospirosis has a high incidence in tropical regions, and has been studied extensively in several Caribbean countries. We studied the carriage of Leptospira serovars by two small mammals which are potential maintenance hosts of the disease in Barbados. A total of 136 mongooses (Herpestes auropunctatus) and 97 mice (Mus musculus) were caught in live traps. Leptospiral antibodies were detected by microscopic agglutination test (MAT) using antigens representing 12 serogroups, and kidney tissues were inoculated into polysorbate medium for isolation of leptospires. The seroprevalence (at a titre of > or = 100) in mice was 28.2% (24/85, 95% CI 19.0, 39.1) and in mongooses 40.7% (48/118, 95% CI 31.7, 50.1). In mice, antibodies were detected predominantly against serogroups Ballum and Autumnalis, while in mongooses the predominant serogroup was Autumnalis. Leptospires were isolated from 28 mice (28.9%, 95% CI 20.1, 39.0) and from 4 mongooses (2.9%, 95% CI 0.8, 7.4). Mouse isolates were identified as serovars arborea (17) and bim (7). As in other parts of the world, common house mice (Mus musculus) represent a significant reservoir of leptospirosis. Although carriage of the Ballum serovar, arborea, was not unexpected, this represents the first time that an animal reservoir of serovar bim has been identified. This is significant because bim causes about 63% of human leptospirosis in Barbados, and control efforts and education for prevention can now be targeted at a specific reservoir.

Animals↗

Virological and serological study in children hospitalized for acute hepatitis (detection of HBsAg, HAAg, anti-HAV and specific IgM-anti-HAV).

Serological methods were used to detect different markers of hepatitis A and B infection in a series of 36 children aged 3.5-13 years with acute viral hepatitis, hospitalized from August 1978 to October 1978. Hepatitis A was verified serologically in 30 patients (83.3%) with the demonstration of specific IgM-anti-HAV. Hepatitis B infection was serologically confirmed in only 2 patients (5.5%). After exclusion of hepatitis A and B as well as of hepatitis due to cytomegalovirus, leptospira and Epstein-Barr virus, 3 patients (8.3) were classified to have post-transfusion non-A non-B hepatitis, and 1 patient (2.5%) showed a sporadic non-A non-B hepatitis. The finding of HBsAg in 5 of 30 patients with hepatitis A suggests that the presence of HBsAg during the acute icteric phase of viral hepatitis is not sufficient for a diagnosis of HB. The presence of HAAg in one of 30 patients with HA confirms previous studies that faecal shedding of HAV stops with the appearance of jaundice. Finally, the above results establish the value of the new radioimmunoassay technique for detection of specific IgM-anti-HAV, which appears to be the best diagnostic test for hepatitis A.

Adolescent↗

[Leptospirosis (author's transl)].

Leptospirosis is one of the most widely distributed zoonoses. Dogs are the pets in which leptospirosis is most common. The use of diagnostic laboratory procedures is discussed. During the first seven days of the disease, leptospires may be detected in the blood and organs; after the seventh day, they are detectable in the kidney and possibly also in the urine. Examination of the serum for antibodies is possible after the first week of the disease. A clinical diagnosis is often hard to establish. Unlike what used to be the case, infections with L. icterohaemorrhagiae are more common today than those with L. canicola. Infections are more common in male dogs than they are in bitches. In addition to antibiotic therapy, preferably consisting in administration of penicillin and streptomycin, symptomatic treatment is useful. Vaccination should be performed in the spring. Dog are probably of minor importance as a source of human infection in the Netherlands.

Animals↗

A cluster of leptospirosis among abattoir workers.

In early December 1998, the Northern Rivers Public Health Unit (north-eastern New South Wales) was alerted to a possible cluster of leptospirosis cases by the supervising scientist of the Western Pacific Region World Health Organization/Food and Agricultural Organization Collaborating Centre for Reference and Research on Leptospirosis. Investigation revealed a cluster of eight leptospirosis cases diagnosed during October and November 1998. All were employees of a local meat works. Leptospira serovars isolated included pomona and hardjo. Symptoms included headache, fever, muscle pain, sore eyes, abdominal pain, vomiting, jaundice, and rash. Five of the eight cases were hospitalised. The infection could not be traced to any particular source. Unfortunately, records of stock killed during the exposure periods were not available. All cases reported exposure to large volumes of animal urine during the course of their work. Protective clothing provided included an apron, gloves, and rubber boots. All of the patients said they wore rubber boots and seven of the eight wore the apron provided. Only two patients reported wearing gloves, the remainder thought these were too difficult to work in.

Abattoirs↗

Immunological reactivity and passive protective activity of monoclonal antibodies against protective antigen (PAg) of Leptospira interrogans serovar lai.

Monoclonal antibody (MAb) AG1 against the protective antigen (PAg) was produced and characterized. It had been extracted from Leptospira interrogans serovar lai by the chloroform-methanol-water method and was of glycolipid nature (23-30Kd). The fact that the PAg was a serogroup-specific antigen was shown by MAb AG1, because MAb AG1 agglutinated serovars of serogroup Icterohaemorrhagiae. Purified MAb LW2 and LW3 which are agglutinating antibodies of serovar lai and AG1 passively protected hamsters from leptospiral infection. Induction of the reactive oxygen intermediates by MAbs from peritoneal exudate macrophages of mice were observed in the chemical luminescence assay and the MAbs reacted with the PAg in the dot enzyme-linked immunosorbent assay. However, MAb LW4a against the genus-specific antigen present in the sub-surface of leptospiral cells did not show protective and reactive-oxygen-inducing activities; they reacted with the non-protective glycolipid antigen of low molecular weight (Fr I, 10-15Kd) in the dot enzyme-linked immunosorbent assay. These results indicated that anti-PAg antibody exhibited opsonic activity against Leptospira and the production of reactive oxygens by macrophages led to leptospiricidal action as one of the defence mechanisms of the host against leptospiral infection. However, the antibody against the genus-specific glycolipid antigen may not be important for protection against leptospiral infection.

Agglutination Tests↗

Antibodies to bovine bacterial and viral pathogens in pronghorns in Alberta, 1983.

Sera from 210 pronghorns (Antilocapra americana) ranging in southeastern Alberta were tested for antibodies to disease agents present in indigenous cattle. No antibodies to Brucella abortus, Leptospira interrogans serovars pomona, hardjo, or grippotyphosa, or infectious bovine rhinotracheitis virus were found. Antibodies at prevalences of 43.8% and 49.2% were detected to bovine virus diarrhea (BVD) and parainfluenza type 3 (PI-3) viruses, respectively. The much higher prevalence of BVD virus antibodies in cattle than in pronghorns, and the occurrence of clinical bovine PI-3 infection in the study area, suggest that cattle may be a source of infection to the pronghorns.

Alberta↗

Seroprevalence of leptospirosis in a rural flood prone district of Bangladesh.

Leptospirosis is a worldwide zoonotic disease. In the present investigation, a total of 89 human sera from a flood prone district of Bangladesh was screened by a one-point microscapsule agglutination test (MCAT). MCAT-positive and -doubtful sera were further tested by microscopic agglutination test (MAT) against 16 reference serovars of Leptospira interrogans, and the antibody titres determined. In MCAT, 34 sera were positive and 22 were doubtful. Among those positive and doubtful sera, 33 and 20, respectively were tested by MAT. Thirty-four out of 53 MCAT-screened samples were MAT-positive. The titres ranged from 20 to 1600 with antibodies to serovars copenhageni, australis, cynopteri and icterohaemorrhagiae being the most prevalent. Eleven MCAT-positive samples failed to react with any strains used by MAT, suggesting the presence of new or untested serovars. Among the MAT-positive samples, the presence of antibody against two or more serovars was more common than that of a single serovar. The present study suggests that rural people in Bangladesh are at high risk to leptospiral infection.

Agglutination Tests↗

Effects of products of autolysis of tissue and urine on the viability, morphology, and antigenicity of Leptospira interrogans serovar pomona.

The effect of the products of autolysis on Leptospira interrogans serovar pomona was investigated in bovine kidney tissues and urine inoculated with this organism. No viable leptospires were found at 24 h or subsequently after inoculation of kidney tissues or urine. Leptospires and their soluble antigens were rapidly destroyed in tissues stored at 20 degrees C and could not be detected by the fluorescent-antibody test and the enzyme-linked immunosorbent assay. On the other hand, leptospires were detectable by dark-field microscopy and fluorescent-antibody test 38 days after addition to urine. The rate of destruction proceeded less rapidly in tissues stored at 4 degrees C. Under the conditions of this study, soluble leptospiral antigen levels decreased less rapidly than did the number of detectable leptospires. Similarly, breakdown of leptospires, demonstrated by dark-field microscopy and fluorescent-antibody test, progressed during the first 5 days after inoculation of urine, when levels of soluble leptospiral antigen remained constant. Although the number of intact leptospires was markedly reduced after freezing of tissues at -20 degrees C, soluble leptospiral antigen levels remained unaltered. Neither intact leptospires nor their soluble antigens were detected after preservation of tissues in Formalin. However, formolization of urine delayed the destruction of leptospires and increased the levels of soluble leptospiral antigen during at least the first 5 days after inoculation. These results indicate that assays such as the enzyme-linked immunosorbent assay should be useful for the detection of soluble leptospiral antigens in urine and in autolyzing tissues such as those taken from a bovine fetus aborted as a result of leptospiral infection.

Animals↗

Leptospires in the marine toad (Bufo marinus) on Barbados.

Leptospires were isolated from the kidneys of four of 211 toads (Bufo marinus) caught on Barbados. Two of the isolates were identified as Leptospira interrogans serovar bim in the Autumnalis serogroup (the most common cause of leptospiral illness on Barbados), and two as possibly new serovars in the Australis serogroup. Sera from 198 of the toads were examined by the leptospire microscopic agglutination test. Forty-two (21%) were positive at titers of greater than or equal to 1:100, and 54 (27%) at greater than or equal to 1:50. The predominating serogroups were Australis (50%), Autumnalis (23%) and Panama (13%). The agglutination tests on the culture-positive toads showed that serologic studies alone may be of limited value in these animals. Bufo marinus can harbor pathogenic leptospires, and it may be a significant source of the Autumnalis serogroup infections in the Caribbean.

Agglutination Tests↗