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Leptospires in Rattus spp. on Barbados.

Among 138 Rattus norvegicus and 98 R. rattus trapped on Barbados in 1964-65 and examined for evidence of leptospiral infection, seropositivity prevalence rates were similar (34 and 30%, respectively), but isolation/dark field microscopy rates were higher in R. norvegicus (27%) than R. rattus (15%). R. norvegicus carried mainly serogroup Icterohaemorrhagiae and R. rattus mainly serogroup Autumnalis. These two serogroups cause 90% of severe human leptospirosis on the island.

Animals↗

[Leptospirosis in children: a not infrequent disease (author's transl)].

A total of 14 cases of leptospirosis in children aged from 5 to 15 years, hospitalized in the Paris region, are reported. The polymorphe character of the disease is pointed out. The necessity for respecting a rigorous chronological order when requesting complementary examinations, which could confirm the diagnosis, is stressed. Domestic animals, team-mates, and while swimming at week-ends or during holidays, are potential sources or means of infection in the young. This justifies searching for leptospirosis in cases of unexplained fever, a pseudo-influenza syndrome, a meningitis which is mainly of the lymphocytic type, or more rarely, in cases of jaundice. Leptospirosis should be a disease well-known by the pediatrician.

Adolescent↗

The effects of the periodical use of in-feed chlortetracycline on the reproductive performance of gilts and sows of a commercial pig farm with a history of clinical and subclinical viral and bacterial infections.

The objective of this study was to assess the effects of in-feed chlortetracycline (CTC) as a measure of preventing or minimizing infectious problems of reproductive failure in gilts and sows. In a farm of 400 Large White x Landrace gilts and sows with a clinical history of porcine respiratory and reproductive syndrome (PRRS) virus, the animals were treated with CTC. Treatment consisted of 10 g CTC sow/day for 15 days every 3 months. It improved the health status of sows by decreasing post-farrowing clinical mastitis and vaginal discharges, abortions, return-to-oestrus and irregular return-to-oestrus rates. These beneficial effects had a positive impact on the performance of the litter. More piglets were born live and weaned. These positive effects improved with repeated use of CTC. The serological evidence of PRRS virus, Leptospira spp. and Chlamydia spp. and the subsequent beneficial use of the antimicrobial agent indicate that reproductive failure, possibly resulting from the bacterial agents can be controlled with in-feed use of broad spectrum antimicrobials.

Abortion, Veterinary↗

[Homology of the recombinant DNA of plasmid pDJH2 with the recombinant DNA probe of L. alstoni and analysis of its expression in Escherichia coli].

Dig-labeled recombinant DNA probe of L. alstoni which contains the entire structural OmpL1 gene was hybridized with the recombinant DNA of the plasmid named pDJH2 of the gene library of L. interrogans serovar lai strain 017. The result showed a high degree of homology among them; expression of recombinant DNA of pDJH2 was achieved by beta-D-galactosidase (IPTG) induction in E. coli. The molecular weight of this product is 68kd. Then they were treated with proteinase K and subjected to SDS-PAGE. The results showed it is a protein in nature. Using the specific monoclonal antibody E4B7D5 on immunoblotting and specific polyclonal antibody on dot-ELISA assay, we investigated the immune reaction and noticed that protein 68kd might be an antigen in character. E. coli which contains the recombinant plasmid pDJH2 were injected into BALB/c mice. Then the mice were challenged by leptospires of the strong virulence strain 017, but all the infected mice survived. In this paper, we first report the expression of recombinant DNA of L. interrogans serovar lai strain 017 in E. coli when injected with IPTG, and immunoprotection of BALB/c mice which were injected with the expression against the infection of L. interrogans serovar lai strain 017. pDJH2 may be the first recombinant for which the gene has been cloned and its expression product 68kd may be the immunoprotective antigens.

Animals↗

Growth, survival, antigenic stability, and virulence of Leptospira interrogans serotype canicola.

Leptospira interrogans serotype canicola (strain NADL A-13) grew from inocula as small as two cells in liquid polysorbate 80 medium (P-80 medium, in P-60, P-40 and P-20 media, and in P-80 medium from which polysorbate, NH4C1 or thiamine had been omitted. It grew well initially in vitamin B12-deleted P-80 medium, but only with inocula as large as 26 x 10(4) cells per ml. P-80 medium lacking both polysorbate and NH4Cl supported light growth from small inocula, but the omission of thiamine and vitamin B12 in addition seriously affected the properties of the medium. Where readily detectable growth did not develop in liquid nutrient-deleted medium, viable ortanisms could often be demonstrated indirectly by subculture to semisolid medium, and their occurrence was influenced by the presence of albumin, thiamine, and vitamin B12. Growth on semisolid media was comparable with that in liquid media of similar composition. The absence of polysorbate 80, thiamine, or vitamin B12 prevented the appearance of Dinger's zones of growth from small inocula. Antigenic composition as measured by microscopic agglutination tests with homologous and heterologous antisera was not appreciably affected by repeated subculturing in various complete and incomplete media. Homogenates of infected-hamster-kidney tissue in bovine serum-albumin diluent still contained viable organisms after 60 days' storage at 23-25 degrees C. Organisms derived from this material after 3 and 16 days' storage showed no loss of virulence. Organisms grown in artificial culture showed no loss of virulence after storage in bovine albumin diluent or phosphate buffer for 7 days at 23-25 degrees C. Cultures of the organism survived without loss of virulence for 15 months in 13 semisolid media of differing complexity. Single colonies derived from five different solid media were grown in semisolid forms of the parent media and stored at 23-25 degrees C for 10 months without loss of virulence.

Animals↗

A waterborne outbreak of leptospirosis.

During the period from July 10-26, 1984, 33 cases of serologically confirmed leptospirosis occurred in a small town in central Italy. The fatality rate, including the deaths of two unconfirmed cases, was 8.6% (3 of 35). Based on serologic evidence, the infection was caused by leptospires of the serogroup Australis. Epidemiologic study showed that the patients contracted the infection by drinking water from a fountain. The source of leptospiral contamination was probably a hedgehog trapped in a reservoir of water not in use but still connected to the water system of the fountain.

Adult↗

Serological survey of selected pathogens of free-ranging foxes in southern Argentina, 1998--2001.

A serological survey was conducted in the Patagonia region of Argentina to estimate the prevalence of nine disease agents within the populations of free-ranging culpeo (Dusicyon culpaeus) and grey (Dusicyon griseus) foxes. The disease agents were Aujeszky's disease virus (ADV), Brucella, canine adenovirus (CAV), canine distemper virus (CDV), canine parvovirus (CPV), Encephalitozoon cuniculi, Leptospira, Neospora caninum, and Toxoplasma gondii. A total of 84 foxes were sampled (28 culpeo and 56 grey), and 73% of the sera had antibodies against one or more pathogens. Among these seropositive sera, 47% of them reacted to only one antigen, while the other 53% reacted to multiple antigens. The presence of antibodies to Toxoplasma (20%), Neospora (44%), Leptospira (30%) and Brucella (18%) suggests that these organisms actively circulate in the area. Antibodies against CDV, CAV and CPV were detected in 2%, 5% and 5% of foxes, respectively. Regarding Encephalitozoon cuniculi and ADV, no evidence of either was found.

Animals↗

Leptospires colonial variations.

Six of 12 relatively freshly isolated and museum strains representing serovars mozdok, monjakov, kazakhstanica I and patoc were heterogeneous in colonia morphology. Recloned colonial variants of one and the same heterogeneous population did not exhibit any differences in antigenic properties in cross reactions of microagglutination and absorption of agglutinins. Along with it among the clones with various colonial types of the two relatively freshly isolated strains of serovar mozdok distinctions in virulence (LD50) for hamsters were marked. Moreover for the clones one of them differences in the level of renal infection (ID50) and in morphology of the cells (hooked and straight) were found. In a solid Tween-80 albumin medium in populations of two colonial variants (serovars monjakov and kazakhstanica I) colonial mutants appeared without any changes in antigenic properties, virulence and cell morphology with frequency of 10-9-10-8 per one bacterium per one generation.

Antigens, Bacterial↗

Isolation of leptospires in diagnosis of leptospirosis.

Though the microscopic agglutination test is commonly used as an aid to diagnosis of leptospirosis, it lacks sensitivity, cannot differentiate vaccinal titers from those of infection, and cannot identify shedders. The ELISA is sensitive but also cannot differentiate titers of vaccination and infection. Leptospire isolation is essential for a final diagnosis. Tissue or fluid samples should be aseptically collected, if possible, and quickly shipped to the diagnostic laboratory in transport medium or on ice, but should not be frozen. Samples to be collected may include urine, milk, kidney, aqueous humor, and fallopian tube or uterine mucosa.

Animals↗

The importance of leptospirosis in Southeast Asia.

The importance of leptospirosis in Southeast Asia was assessed in conjunction with other studies supported by the U.S. Naval Medical Research Unit No. 2 (US NAMRU-2), Jakarta, Republic of Indonesia. These included studies of hospital-based, acute clinical jaundice in Indonesia, Lao PDR, and Socialist Republic of Vietnam; nonmalarial fever in Indonesia; and hemorrhagic fever in Cambodia. Background prevalence estimates of leptospiral infection were obtained by a cross-sectional, community-based study in Lao PDR. Laboratory testing methods involved serology, microscopic agglutination test, and reverse-transcriptase polymerase chain reaction. Suggestive evidence of recent leptospiral infections was detected in 17%, 13%, and 3% of patients selected on the basis of non-hepatitis A through E jaundice, nonmalarial fever, and hemorrhagic fever (in the absence of acute, dengue viral infections). Leptospiral IgG antibody, reflective of prior infections, was detected in 37% of human sera, collected in Lao PDR. The predominant leptospiral serogroups identified from cases with clinical jaundice were Hurstbridge, Bataviae, and Icterohaemorrhagiae tonkini LT 96 69. Among the nonmalarial febrile cases, Bataviae was the most frequently recognized serogroup. Pyrogenes and Hurstbridge were the principal serogroups among the hemorrhagic fever case subjects. These findings further attest to the relative importance of clinical leptospirosis in Southeast Asia. The wide spectrum of clinical signs and symptoms associated with probable, acute, leptospiral infections contributes to the potential of significant underreporting.

Adolescent↗

Use of furosemide to obtain bovine urine samples for leptospiral isolation.

The use of the diuretic furosemide made it possible to obtain samples of urine from cattle for leptospiral isolations. The drug was injected IV at a dose level of 0.8 mg/kg for heifers and 0.5 mg/kg for calves. The average time to first voiding in heifers was 19 minutes. The average time from the first to the second voiding was 17 minutes. The average time to the first voiding in four calves was 12 minutes; the average time from the first to the second voiding was 10 minutes. A decrease in urinary osmolarity provoked by furosemide created a more favorable condition for the survival of leptospires. Leptospires were isolated in 24 (72.7%) of 33 weekly cultural attempts with the aid of furosemide in three experimentally infected adult cattle. Serovar hardjo was isolated in 16 (57.1%) of 28 weekly cultural attempts with aid of the diuretic in four experimentally infected calves. The recovery frequency was 28.5% from the first voiding and 50% from the second. Leptospires were not isolated from urine obtained from the calves by manual stimulation. Untoward side effects that might have been attributable to furosemide were not observed. Furosemide appears to be well suited to obtain urine samples from cattle for leptospiral isolation.

Animals↗

Enzyme-linked immunosorbent assay and immunoblot analysis for detection of antibodies to Borrelia burgdorferi in dogs. The impact of serum absorption with homologous and heterologous bacteriae.

Sera from 665 apparently healthy dogs were examined for antibodies to the Lyme disease spirochete Borrelia (B.) burgdorferi by using an ELISA with a whole cell sonicate of B. burgdorferi sensu stricto reference strain B31 (ATCC 35210) as antigen. To discover false positive reactions due to the unsatisfactory specificity of conventional enzyme-linked immunosorbent assays for B. burgdorferi, sera were absorbed in parallel with both B. burgdorferi and a heterologous sorbent consisting of whole cells of Escherichia coli, Salmonella typhimurium, and eight serovars of Leptospira interrogans. The difference between optical densities obtained in the ELISA after serum absorption with the heterologous sorbent and the B. burgdorferi sorbent was defined as a new value, "ODdiff", for ELISA reactivity specific for B. burgdorferi. ELISA results were confirmed by immunoblot studies. By testing unabsorbed sera, 48 of 665 serum samples (7.2%) were considered ELISA positive. 37 of these 48 sera (77.1%) were apparently false positive: here a similar reduction of ELISA reactivity was obtained after absorption with B. burgdorferi antigen and with the heterologous sorbent (ODdiff approximately equal to 0). None of these 37 sera gave immunoblot patterns characteristic for canine B. burgdorferi infection.

Animals↗

Development of a polymerase chain reaction test to confirm Mycobacterium avium subsp. paratuberculosis in culture.

A polymerase chain reaction (PCR) assay for confirmation of Mycobacterium avium subsp. paratuberculosis was developed using the primer set derived from ISMav2. The PCR product was 494 base pairs (bp) and could be digested with ClaI, which produced 311- and 183-bp fragments. No amplification of 494-bp DNA fragment was detected from DNA of other Mycobacterium spp., including Mycobacterium avium complex, other bacteria, including Escherichia coli, Pseudomonas aeruginosa, Actinobacillus pleuropneumoniae, Leptospira interrogans serovar pomona, Corynebacterium pseudotuberculosis, Salmonella typhimurium, Borrelia burgdorferi, and Staphylococcus aureus, and the Scedosporium sp. This PCR assay could detect 5-8 genome equivalents.

Animals↗

Detection of leptospires in urine by PCR for early diagnosis of leptospirosis.

We tested urine samples from patients at different stages of current leptospirosis and thereafter to determine whether use of the PCR for detection of leptospires in urine can be a valuable alternative to culturing. The procedure of DNA extraction and subsequent PCR applied to 15 freshly voided urine samples proved to be twice as sensitive as culturing. Overall, we were able to detect leptospires in approximately 90% (26 of 29) of the urine samples. Urine and serum samples were obtained from seven patients, before the eighth day of illness. Although it is generally assumed that leptospiruria starts approximately in the second week of illness, we were able to detect leptospires in all of these early urine samples. In contrast, only two of seven corresponding serum samples gave positive PCR results, which suggests that PCR analysis of urine can be more successful for early diagnosis of leptospirosis than PCR analysis of serum. Urine samples from six patients who had been treated with antibiotics at the time of illness were positive by PCR, implying that the patients were still shedding leptospires in their urine despite treatment. Some of these samples were even taken years after the infection, indicating that shedding of leptospires in urine may last much longer than is generally assumed. We conclude that detection of leptospires in urine with PCR is a promising approach for early diagnosis of leptospirosis and may also be useful in studying long-term shedding.

Acute Disease↗

Pulmonary leptospirosis.

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.

Female↗

Detection of leptospiral DNA by nucleic acid hybridisation with 32P- and biotin-labelled probes.

Dot hybridisation with 32P- and biotin-labelled probes prepared from leptospiral DNA was performed to develop a sensitive and specific diagnostic method for early infection with leptospires. The smallest amounts of leptospiral DNA that could be detected with 32P- and biotin-labelled probes was 1.5 pg and 5 pg, respectively, corresponding to about 750 and 2500 leptospires. Dot hybridisation with a 32P-labelled probe detected leptospiral DNA in sera from all of 14 experimentally infected golden hamsters. The smallest amount of leptospiral DNA detected in these experiments corresponded to about 2500 leptospires. In the test conditions described in this study, the sensitivity of dot hybridisation with a biotin-labelled probe was lower. Little cross-hybridisation was observed with unrelated DNAs which indicates that dot hybridisation could be a useful diagnostic method.

Animals↗

Clinical distinction and evaluation of leptospirosis in Taiwan--a case-control study.

BACKGROUND: Leptospirosis is the most widespread zoonosis. Leptospirosis remains underreported in Taiwan because of ignorance and the broad spectrum of clinical manifestations. Acute renal failure (ARF) is a prominent feature of leptospirosis. This investigation conducted a case-control study to obtain information to distinguish leptospirosis from other conditions with similar presentations. METHODS: Leptospirosis surveillance was performed at Chang Gung Memorial Hospital, Taiwan, between September 2000 and December 2001. Suspected clinical cases were included in the sample and investigated. Diagnosis was confirmed with four-fold or greater increase of microscopic agglutination test (MAT) titer in paired sera; positive immunoglobulin M (IgM) dipstick with single MAT > or =400; or isolation of leptospira. Cases were classified as excluded based on confirmed etiology other than leptospirosis or negative paired serologic test. RESULTS: Twenty-two confirmed cases and 21 excluded cases of leptospirosis were identified from among 169 suspected cases. An outbreak was observed during the flooding from Typhoon Nali. L. shermani, the most common serovar in Taiwan, was identified in 78.5% of confirmed cases. In the confirmed group, mean age was 44 +/- 14.5 (21-66) yrs similar to that of the excluded group, with male predominance (86.4 vs. 57.1%, p<0.05). The most common presentations in the confirmed group were fever (95.5%), ARF (86.4%), myalgia (72.7%) and jaundice (63.6%). Ten patients were infected through occupational or recreational exposure, and in six patients, the infection was associated with flooding. The most distinct presentations of leptospirosis cases compared with excluded cases were increased incidence of hemorrhagic diathesis (odds ratio (OR): 10, p=0.04), myalgia (OR: 8.0, p=0.02), bilateral enlarged kidneys (OR: 7.5, p=0.0004), risk factor exposure (OR: 6.9, p=0.005), sterile pyuria (OR: 6.3, p=0.017), hypokalemia (OR: 5.0, p=0.035) and thrombocytopenia (OR: 4.8, p=0.04). Hospitalization days correlated well with levels of peak creatinine (Cr) (p=0.0362) and platelet nadir (p=0.0039) reached. Penicillin treatment was followed by rapid symptoms and renal function improvement. CONCLUSIONS: Prompt recognition of the characteristic presentations of leptospirosis, followed by timely antibiotic treatment, can dramatically save the patients even with severe multiple organ damage.

Adult↗