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Identification and analysis of genes present in Leptospira interrogans serovar lai but absent in L. biflexa serovar monvalerio.

Genes present in virulent bacterial strains but absent in avirulent close relatives can be of great biologic and clinical interest. This project aimed to identify strain specific DNA sequences of Leptospira interrogans serovar lai, which is absent in the saprophytic L. biflexa serovar monvalerio, via suppression subtractive hybridization with the former as the tester while the latter as the driver. The mixture of PCR amplified DNA fragments from two subtractive hybridization experiments were cloned into pMD18-T vector and the positive clones were identified by dot blotting against the chromosome DNA of the two strains individually. After DNA sequencing and analysis, the distribution of these genomic fragment sequences in a panel of pathogenic and nonpathogenic leptospires was investigated employing dot blot analysis. Among the 188 positive clones randomly chosen, 24 contained the tester strain specific genomic regions, of which, 5 were non-coding fragments while the others contained 23 distinct protein coding sequences. Besides 9 genes encoding functional proteins, 12 genes encode unknown proteins and the rest two genes encode proteins with recognizable domain structures, one for a putative leucine-rich repeats (LRR) family protein while the other as an outer-membrane protein. Our experiment results indicated that suppression subtractive hybridization is effective for screening specific DNA sequences between two leptospiral strains, and some of these sequences might be responsible for virulence determination. Further analysis of these DNA sequences will provide important information on the pathogenesis of Leptospira.

DNA, Bacterial↗

Leptospirosis-induced meningitis and acute renal failure in a 19-month-old male child.

An unusual case of leptospirosis is described in a 19-month-old male child presenting with meningitis and acute renal failure without jaundice. Some aspects concerning the pathogenesis and treatment of this potentially life-threatening disease are also discussed. Leptospirosis was diagnosed on the basis of history and serological tests.

Acute Kidney Injury↗

Leptospiral infection in aborted equine foetuses.

During an investigation of equine abortion, leptospiral infection was demonstrated in nine out of 22 foetuses examined by immunofluorescence and culture. Strains belonging to four serogroups (Australis, Pomona, Hebdomadis and Icterohaemorrhagiae) were isolated. The age of leptospira infected foetuses ranged from six months to term.

Abortion, Veterinary↗

Serological survey for selected diseases in the endangered San Joaquin kit fox (Vulpes macrotis mutica).

Blood from endangered San Joaquin kit foxes (Vulpes macrotis mutica) inhabiting the Elk Hills Naval Petroleum Reserve, Kern County, and the Elkhorn Plain, San Luis Obispo County, California, was collected in 1981, 1982 and 1984 and sera were tested for antibodies against 10 selected pathogens. Proportions of kit fox sera containing antibodies against pathogens were: canine parvovirus, 100% in 1981-1982 and 67% in 1984; infectious canine hepatitis virus, 6% in 1981-1982 and 21% in 1984; canine distemper virus, none in 1981-1982 and 14% in 1984; Francisella tularensis, 8% in 1981-1982 and 31% in 1984; Brucella abortus, 8% in 1981-1982 and 3% in 1984; Brucella canis, 14% in 1981-1982 and none in 1984; Toxoplasma gondii, 6% in 1981-1982; Coccidioides immitis, 3% in 1981-1982; and Yersinia pestis and Leptospira interrogans serotypes canicola, grippotyphosa, hardjo, icterohaemorrhagiae, and pomona, none in 1981-1982. Although antibodies against selected pathogens were present, no clinical indications of disease were observed in these fox populations.

Animals↗

Serologic survey for selected microbial pathogens in bison from Kansas.

A serologic survey was conducted on an American bison (Bison bison) herd in Kansas for antibodies against Brucella spp., Leptospira interrogans serovar canicola, pomona, grippotyphosa, icterohaemorrhagiae, and hardjo, Anaplasma spp., bluetongue virus, infectious bovine rhinotracheitis virus and bovine viral diarrhea virus. There was an increase in prevalence of bluetongue antibodies from 38% in 1987 to 100% in 1989 in animals greater than or equal to 24-mo-old. Prevalences of antibodies against the other livestock pathogens were either negative or at levels associated with previous vaccination.

Anaplasmosis↗

[A case of acute pancreatitis due to Weil's disease].

Leptospirosis, the most common vasculitic zoonosis in the world, is characterized with jaundice and acute renal failure. However, pancreatitis is an uncommon complication of leptospirosis. In this report, an acute pancreatitis case due to Weil's disease has been presented. A 31-year-old female patient with high levels of glucose, blood urea nitrogen, creatinine, creatine kinase, bilirubin, amylase and lipase, has been diagnosed to have leptospirosis by the high positive result (1/800) of microscopic agglutination test against Leptospira interrogans serogroup icterohemorrhagiae. The patient has been treated with supportive and symptomatic therapy, and with penicillin G for leptospirosis. Following triple hemodialysis, all the blood biochemistry tests returned to normal on the tenth day of therapy. This case was reported to draw attention to Leptospira infections which should be considered in the differential diagnosis of patients with jaundice and pancreatitis.

Acute Disease↗

[Clinical reasoning and decision-making in practice. A man with jaundice, renal function disorder, thrombocytopenia, muscular pain and confusion].

A 51-year-old man became jaundiced, after experiencing flu-like symptoms for a week. On admission anaemia, thrombocytopenia, and hepatic and renal function disorders were present. One month previously he had had sinusitis. Thrombotic thrombocytopenic purpura (TTP)/haemolitic-uraemic syndrome (HUS) was suspected, but the thrombocyte count rose spontaneously. The leukocytosis indicated an infectious cause of the symptoms and the jaundice appeared to originate from intrahepatic cholestasis. All symptoms could be traced to leptospirosis icterohaemorrhagica (known as 'Weil's disease'). The source was possibly the patient's fishpond. He recovered spontaneously and was also treated with antibiotics. Leptospirosis, a multi-organ disease is seen ever more infrequently as an occupational disease in stock farmers and increasingly frequently in people whose hobbies involved animals.

Anti-Bacterial Agents↗

[Screening for antibodies against zoonotic agents among employees of the Zoological Garden of Vienna, Schönbrunn, Austria].

The aim of this study was to investigate the prevalence of antibodies against zoonotic agents in employees of the zoological garden of Vienna, Schönbrunn, Austria. Sixty out of 120 employees participated in the study. In 97% of them antibodies to at least one zoonotic agent were identified. Only two participants were free of antibodies to the zoonotic agents tested. The following seroprevalences (in brackets) were obtained: Viral zoonotic (and potentially zoonotic) agents: Influenzavirus A/H1N1 (58%), Influenzavirus A/H3N2 (85%), Lymphocytic choriomeningitis virus (13%), Encephalomyocarditis virus (5%), Orthopox- (Cowpox-) virus and Hantavirus type Puumala (3%). Hantavirus type Hantaan and Borna disease virus (all negative). Bacterial zoonotic agents: Bartonella henselae (65 %), Borrelia burgdorferi (10%), Leptospira interrogans serovar copenhageni and serovar icterohaemorrhagiae as well as Chlamydophila psittoci (2% each). Brucella spp., Coxiella bumetii, and Francisella tularensis (all negative). Parasitic zoonotic agents: Toxoplasma gondii (53%), Toxocara spp. (21%), Capillaria hepatica (2%), Fasciola hepatica, Schistosoma mansoni, E. multilocularis, and E. granulosus (all negative). The remarkably high seroprevalence to the causative agent of cat scratch disease, Bartonella henselae, is probably due to the private contact of the employees to cats. Regarding viral zoonotic agents it has to be mentioned that Influenzavirus vaccination and/or human-to-human transmission of especially A/H3N2 Influenzaviruses probably attributed significantly to the very high seroprevalence to both Influenzavirus types A/H1N1 and A/H3N2. When investigating parasitic zoonotic agents, high prevalence rates were found against Toxoplasma gondii and Toxocara spp., however, it was not possible to establish a causal link between seropositivity and the professional activity in the zoo. Interestingly, in the case of antibodies to T. gondii, the typical correlation with age was not found in this study, while in the case of the Toxocara spp. positive subjects a correlation was identified with both age and duration of employment in the zoo. Regarding the later two zoonotic parasites, employees of the zoological garden showed significantly higher seroprevalences than the average Austrian population. Antibodies to Capillaria hepatica, a hepatic-parasite in rodents which is diagnosed in humans rarely, were identified in one employee and another one showed a questionable positive result. Further investigations did not exhibit clinical infestations with the parasite in these two individuals so far.

Adolescent↗

Prevention of renal carriage of leptospirosis in dogs by vaccination.

Dogs vaccinated with a Leptospira interrogans vaccine containing serogroups Canicola and Icterohaemorrhagiae and prepared from cultures grown in a protein-free medium resisted challenge with heterologous representatives of these serogroups. In contrast, control dogs were pyrexic and leptospires were isolated from the blood for nine days following Canicola challenge and six days after Icterohaemorrhagiae challenge. Leptospires were isolated from the urine of controls throughout the post challenge period and from kidneys at post mortem examination of six out of six and four out of six Canicola and Icterohaemorrhagiae challenged controls, respectively. There have been no reports of hypersensitivity reactions to this vaccine since its introduction in 1980.

Animals↗

Classification of Leptospira interrogans serovar lai strain 017 by using monoclonal antibodies.

Leptospira interrogans serovar lai was identified in China in 1966 as a new serovar of the icterohaemorrhagiae serogroup by cross-absorption tests. In this study, we established three hybridoma cell lines producing monoclonal antibodies (MAbs) of the immunoglobulin G3 subclass (LW1, LW2, and LW3) and of the immunoglobulin M class (LW4a) against serovar lai strain 017 by the cell fusion technique. Immunological reactivities of the MAbs were determined by the microscopic agglutination test, enzyme-linked immunosorbent assay (ELISA), and indirect immunofluorescence assay. MAbs LW1 and LW3 agglutinated cells of serovars lai, birkini, and gem of serogroup icterohaemorrhagiae. LW2 agglutinated various serovars of serogroup icterohaemorrhagiae, except for serovar tonkini. LW4a reacted positively with various Leptospira species, including a new species, Leptospira parva, in the ELISA and indirect immunofluorescence assay. However, LW4a did not react with Leptonema illini 3055. The results of an inhibition ELISA with heated outer envelope (OE) or periodate-oxidized OE suggested that these MAbs recognize a carbohydrate moiety of the OE as the antigenic determinant.

Antibodies, Monoclonal↗

[Formation of group and intergroup antibodies in humans with leptospirosis infections].

The authors studied regularities attending the biosynthesis of group (homologous) and intergroup (heterologous) antibodies in man during the whole cycle of leptospirosis infection, and for three months after it. Leptospira of the serological groups Pomona, Grippotyphosa, Icterohaemorrhagiae, Hebdomadis, the most prominent in the morbidity structure at present, served as etiological causative agents of the disease. Biosynthesis of both homologous and heterologous antibodies had its specific features for each serological group of the disease, and was characterized by the quadripol dynamics of 19S-macro and 7S-microglobulin antibodies production in leptospiroses of serological groups Pomona and Gryppotyphosa, whereas in leptospiroses of serological groups Icterohaemorrhagiae and Hebdomadis a graphically dipolic synthesis dynamics of 19S-macroglobulin antibodies alone was noted.

Antibody Formation↗

Serologic studies on brucellosis, leptospirosis and tularemia in moose (Alces alces) in Quebec.

Blood samples were obtained from 208 moose in La Vérendrye and Matane Reserves and in Laurentides Park, Quebec, Canada. Sera were tested for antibodies to Brucella abortus, Leptospira interrogans serovar ballum, canicola, grippotyphosa, hardjo, icterohaemorrhagiae and pomona, and Francisella tularensis. Fifteen sera contained evidence of prior exposure to F. tularensis. Only one animal was a seroreactor to L. interrogans serovar grippotyphosa and none of them had antibodies to B. abortus.

Animals↗

Isolation of Icterohaemorrhagiae from Microtus monte belli trapped at the Nagaoka area, Niigata Prefecture in Japan.

Three strains of leptospira were isolated from field rodents on the occasion of scrub typhus survey on the Nagaoka, Kurojo, Kamikawanishi area, Niigata Prefecture in September 1961. Out of 10 Microtus montebelli 3 strains of leptospira by means of heart puncture in febrile stage from guinea pigs. They were identified as icterohaemorrhagiae by cross-agglutination-lysis and a cross absorption tests. It was the first report of icterohaemorrhagiae in Microtus montebelli in Japan.

Animals↗

Epidemiological studies on leptospirosis in Chiang Mai (Thailand).

A total of 270 serum samples collected in Chiang Mai province were examined for antibodies against leptospira using the microscopic agglutination test (MAT). Four of 40 serum specimens from patients who visited the hospital with the common cold, were positive with a titre of 20. Twelve (10.4%) of the 115 samples in the Doi Saket district showed a positive reaction. Only 2 of 115 sera of school children in Chiang Mai city had antibodies. Specific serovars detected were Leptospira hebdomadis (5), L. australis (3), L. icterohaemorrhagiae (2), L. bataviae (2), and one each of L. canicola, L. javanica and L. pyrogenes. One case of mixed infection with L. hebdomadis and L. javanica, and L. autumnalis and L. australis were observed.

Adolescent↗