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Human leptospirosis in Italy, 1986-1993.

In the eight-year period 1986-1993, the Italian National Center for Leptospirosis and the Regional Leptospira Laboratories confirmed 312 cases of clinical leptospirosis by using the microscopic agglutination (MA) assay. The majority of cases was observed in Northern regions of the Country. Cases were reported in all age groups, but were most common in the working-age population. Of 312 cases, 291 (93.3%) occurred among males. The largest number of infections was ascribed to occupational activities (45.8%). The typical leptospiral seasonal course, with a peak during the summer, was observed. Involvement of the liver was the most frequent manifestation. Influenza-like symptoms were the only signs of illness in 11.1% of cases. Anti-leptospira antibodies, cross-reacting with two or more serovars, were found in 28.2% of sera. The most frequent serovar-specific antibodies were those against poi, icterohaemorrhagiae, bratislava, copenhageni and sejroe.

Adolescent↗

Interactions of virulent and avirulent leptospires with primary cultures of renal epithelial cells.

A primary culture system for the cells of mouse renal-tubular epithelium was established and used to observe the adhesion of leptospires. Virulent strains of serovars copenhageni and ballum attached themselves to epithelial cells within 3 h of infection whereas an avirulent variant of serovar copenhageni did not adhere to epithelial cells at all within the experimental period of 24 h. The saprophytic Leptospira biflexa serovar patoc became attached non-specifically to inert glass surfaces as well as to the cells. The adhesion of leptospires to epithelial cells was not inhibited by homologous antibody.

Adhesiveness↗

Effect of heat or chemical treatment on leptospiral antigens.

The use of bacterins is one of the primary methods in the control of leptospirosis in domestic animals, especially cattle, swine, and dogs. Bacterins have been made using chemically treated or heat-inactivated organisms. Many of the initial studies indicated that the bacterins gave complete protection, whereas later reports found that some inactivation methods resulted in bacterins that protect against death but not against kidney infection and renal shedding. This raised the possibility that some of the antigens had been altered. To investigate this possibility, we heat or chemically treated Leptospira interrogans serovar icterohaemorrhagiae CF-1, L. interrogans serovar canicola Moulton, L. interrogans serovar grippotyphosa SC4397, L. interrogans serovar hardjo Hardjoprajitno, and L. interrogans serovar pomona MLS. Temperatures used to determine alterations in antigens were 50, 56, 80, 100, and 121 degrees C. The chemicals used were formaldehyde, phenol, and thimerosal. Analysis was done using Laemmli gels and Western blots. We found that heating at 50 or 56 degrees C had the smallest effect on antigens, whereas heating at 80 or 100 degrees C caused the appearance of high-molecular-weight bands on Western blots. Heating at 121 degrees C caused almost all the bands to disappear, in both the stained gel and the Western blot. Both phenol and Formalin treatment altered proteins, whereas thimerosal treatment appeared to have little effect compared with heating at 50 degrees C.

Antigens, Bacterial↗

[A small-mammal study in the northeastern Kara Kum for leptospirosis infectivity].

Search for natural foci of leptospirosis was carried out in 1987-1989 in humid biotopes of Tashauz Province, Turkmenistan. Such potential carriers of leptospirosis as house mice (Mus musculus) and tamarisk gerbils (Meriones tamariscinus) are widely spread in this area, and the size of their population can amount to great numbers. The sharpest fluctuations in the population size are characteristic of house mice inhabiting the shores of water collecting ponds and lakes in the regions of minimally used pastures. A moderate number of house mice was registered in spring and autumn at the area of irrigated agriculture. Only here and only in autumn a faint epizootic manifestation of the natural focus of L. grippotyphosa infection in house mice was registered for the first time in Turkmenistan. The titers in mouse blood sera, determined in the microagglutination test and the Leptospira lysis test, were 1:20 to 1:200. According to the data on the humidity and pH of the soil in the vicinity of irrigated fields, spring and summer months may be regarded as the most favorable period for the survival of leptospires in the environment. The probability of the aggravation of the epizootic situation seems to increase when irrigated fields adjoin pastures of are temporarily used as grazing ground for cattle.

Agriculture↗

Polymerase chain reaction for the laboratory diagnosis of aseptic meningitis and encephalitis.

A protocol for testing cerebrospinal fluid specimens using a range of PCR assays for the diagnosis of central nervous system infection was developed and used to test prospectively 383 specimens. PCR assays were used for the detection of adenovirus, Borrelia burgdorferi, enteroviruses, Epstein Barr virus, cytomegalovirus, herpes simplex virus, human herpes virus type 6, JC virus, Leptospira interrogans, Listeria monocytogenes, lymphocytic choriomeningitis virus, measles virus, mumps virus, Mycobacterium sp. , Mycoplasma pneumoniae, Toxoplasma gondii and varicella zoster virus. Of the 383 specimens tested in this study, 46 (12.0%) were found to be positive. The microorganisms detected were CMV, enterovirus, Epstein Barr virus, herpes simplex virus, human herpes virus type 6, JC virus, L. monocytogenes, Mycobacterium genus, Toxoplasma gondii and varicella zoster virus. The introduction of the PCR protocol described has improved the diagnosis of a range of central nervous system infections in our laboratory. We believe however that further evaluation of these assays in immunocompromised patients is necessary to better determine the predictive value of positive PCR results in these patient groups.

Adolescent↗

The susceptibility of a strain of Leptospira interrogans serogroup icterohaemorrhagiae to amoxycillin, erythromycin, lincomycin, tetracycline, oxytetracycline and minocycline.

The failure of prophylactic penicillin to prevent a laboratory acquired case of Icterohaemorrhagiae leptospirosis prompted determination of the MIC and MBC of amoxycillin, erythromycin, lincomycin, tetracycline, oxytetracycline and minocycline for the infecting strain. Amoxycillin followed by erythromycin were the most effective, with MBCs of 0.5 mg/l after 7 days exposure and 0.1 mg/l after 21 days exposure respectively. Leptospires grew in the presence of high concentrations of tetracycline hydrochloride and oxytetracycline after prolonged incubation. This effect was less pronounced with minocycline, with MIC's of 0.025, 0.05 and 0.1 mg/l after 7, 14 and 21 days exposure respectively. The MIC of lincomycin was 0.25 mg/l at each time interval. These results support the high dose, long duration antibiotic regimens recommended in the literature.

Amoxicillin↗

Prevalence of Leptospira interrogans serovar hardjo in the genital and urinary tracts of non-pregnant cattle.

In a bacteriological investigation of 60 cows and heifers from an abattoir Leptospira interrogans serovar hardjo was isolated from 39 (65 per cent) animals. The genital tract (57 per cent) appeared to be as important a site of hardjo localisation as the urinary tract (62 per cent). This is believed to be the first report of genital hardjo infection in naturally infected cattle.

Abattoirs↗

The persistence of leptospiral agglutinins titers in human sera diagnosed by the microscopic agglutination test.

The persistence of agglutinins detected by MAT has created some problems to the interpretation of the results. The aim of this study was to examine the data of serology from 70 patients with serologically confirmed diagnosis of leptospirosis by during 3-13 months after being affected with leptospires in order to elucidate the interpretation of the persistence of agglutinins detected by MAT. Sixty-one patients sera (87.14%) had titers equal or greater than 800. Of these, two individuals maintained titers of 800 thirteen months after the onset. This study showed that only one sample of sera with high titers is not reliable to determine the time at which infection occurred.

Agglutination Tests↗

Deoxynucleotide sequence conservation of the endoflagellin subunit protein gene, flaB, within the genus Leptospira.

A Polymerase chain reaction was developed to amplify the entire open reading frame of flaB, the gene encoding the endoflagellin subunit protein. The 852 bp amplified products from 23 serovars of the genus Leptospira were subjected to restriction endonuclease analysis and the profiles correlated well with phylogenetic relationships between these serovars. The flaB deoxynucleotide sequences of L. hardjo-bovis, L. hardjo-prajitno and L. grippotyphosa were determined. The deduced primary amino acid sequences of each were highly conserved with only three amino acid residue differences observed. The deoxynucleotide sequences showed genetic drift with alternative bases in the third position of codons. The PCR product derived by amplification of flaB from L. grippotyphosa was cloned into the expression vector pGEX-2T and a recombinant FlaB fusion protein made. As predicted from the deduced amino acid sequences, the recombinant FlaB cross-reacted with heterologous antiserum derived from a rabbit infected with L. hardjo-bovis.

Amino Acid Sequence↗

Laboratory and clinical features of experimental feline leptospirosis.

In order to assess the clinical, laboratorial and epidemiological aspects of feline leptospirosis, ten female and male adult cats were experimentally inoculated with pathogenic and autochthonous field isolate of Leptospira interrogans. Five of them were inoculated subcutaneously with serovar icterohaemorrhagiae (R-192) and the others five with serovar canicola. No clinical and laboratorial alterations were found in these animals. Antileptospiral agglutinins were detected in 90% of the infected cats, shortly after the 1st week after inoculation. The leptospiral agglutinins were detected for 8 to 12 weeks and the elimination of leptospires through urine was observed only in animals infected with serovar canicola, beginning 2 to 4 weeks after inoculation and lasting for 2 to 8 weeks. Isolations of leptospires from blood and kidneys were unsuccessful.

Animals↗

Isolation of Leptospira noguchii from sheep.

The main goal of this study was to obtain new isolates of Leptospira spp. from sheep. A total of 10 kidney samples and 44 blood samples were collected from sheep slaughtered in Pelotas, Southern Brazil. One isolate was obtained which was identified by 16S rRNA gene sequencing and serogrouping to be Leptospira noguchii serogroup Autumnalis. Microscopic agglutination test (MAT) evaluation revealed that 4.5% of the sheep sera reacted against the Autumnalis serogroup. This is the first report of isolation of L. noguchii from sheep. Together these findings indicate that L. noguchii infections may be a potentially important veterinary problem in this domestic animal species.

Agglutination Tests↗

Human leptospirosis in Erode, South India: serology, isolation, and characterization of the isolates by randomly amplified polymorphic DNA (RAPD) fingerprinting.

The study describes the first attempt to record leptospirosis in Erode by isolation and serological tests such as the microscopic agglutination test and IgM-based enzyme-linked immunosorbent assay. Twenty-nine clinically suspected cases showing fever, headache, body ache associated with jaundice, decreased urine output, and conjunctival suffusion were included. The age of the patients ranged between 10-71 years and most of them were agricultural workers. Paired sera were possible among 12 cases. All the patients had fever and headache and other more common symptoms were myalgia and icterus. Leptospiral culture was positive in 7 (24.1%) patients. Out of 29 patients, 26 (89.7%) were diagnosed as having current leptospiral infection based on serology and isolation. The leptospiral isolates KSR 1-6 were further characterized by using randomly amplified polymorphic DNA fingerprinting shown genetic similarities with Leptospira interrogans spp. This study shows the presence of leptospirosis among the hospital cases of Erode and that this disease is a potential health hazard of agricultural workers in Cauvery basin.

Adolescent↗