Human Hardjo leptospirosis detected in the Slovak Republic by using serum antibody absorption test.

OBJECTIVE: The study was aimed at the discovery of the possible presence of the human Hardjo leptospirosis in Slovakia. BACKGROUND: Throughout the world, there is a considerable number of publications confirming the presence of this zoonotic infection in domestic animals. On the contrary, scarcely more than a dozen of reports dealing with this occupational disease in humans was published till now. Moreover, the results of them are not convincing in all cases. MATERIAL AND METHODS: Patients´ and dairy cattle serum samples with detected antibodies against the Leptospira strains belonging to four different serovars of the serogroup Sejroe known to be present in Slovakia (i. e. Sejroe, Istrica, Saxkoebing, Polonica) and also reacting with serovar Hardjo strains were examined by using the serum-agglutinin absorption tests in order to determine the etiological agent of the illness on serovar level.  RESULTS: The Hardjo infection was unequivocally confirmed approximately in 1/5 of patients´sera and in 3/5 of dairy cattle sera. CONCLUSION: The AATs are useful tools for detection of Hardjo leptospirosis what is a necessary condition for determination of appropriate epidemiological measures related to this occupational disease. As our knowledge, this methodological approach was not used by other authors till now (Tab. 4, Fig. 1, Ref. 19).

Human Mozdok leptospirosis first diagnosed by serum agglutinin-absorption tests in the Slovak Republic.

INTRODUCTION: The serovar Mozdok related leptospirosis in humans were not yet feasibly diagnosed using merely the standard micro-agglutination test (MAT) what was perhaps due to the impossibility to distinguish them from illnesses that are caused by Leptospira strains belonging to other serovars of the serogroup Pomona. On the contrary, leptospires of the Mozdok serovar were cultured from rodents and domestic animals world-wide including Central Europe where only Leptospira strains of the serovars Pomona and Mozdok are known to be present till now. STUDY OBJECTIVE: The aim of the study was to discover if leptospires of Mozdok serovar may cause human leptospirosis that remained hidden till now among infections diagnosed merely by MAT as Pomona illnesses. MATERIAL AND METHODS: The reference Leptospira strains of Pomona and Mozdok serovars (Pomona and 5621), as well as three endemic, and in some tests only two strains of human and pig origin (Simon, S-23, Pöstényi), and two strains of rodent provenance - Apodemus agrarius (M-210/98 and M-71/01) were used for this purpose. First, the endemic strains were assigned to one of the afore-mentioned two serovars by agglutinin cross-absorption tests performed using rabbit immune sera, monoclonal antibodies and random amplified polymorphic DNA methods. Afterwards, twenty-one sera of patients with a Pomona leptospirosis confirmed by MAT were examined by agglutinin absorption test (AAT). RESULTS: Based on the results of the mentioned laboratory method used, the endemic Leptospira strains of human and pig origin could be affiliated to the serovar Pomona, while those of rodent origin were classified as serovar Mozdok strains. Out of the 21 patients sera, an illness caused by the serovar Mozdok strains was found out in 13 cases and a disease caused by serovar Pomona strains in 8 cases. Their differentiation was made on the strength of the following results of AATs: All strains from the serovar Mozdok have completely absorbed antibodies (anti-Pomona and anti-Mozdok) from the tested sera, however following the absorption of these sera with the Pomona strains, high levels of residual antibodies reacting in MAT with the Mozdok strains have still persisted. In this way, it was possible to prove the Mozdok infection in thirteen patients. On the contrary, following the absorption of the sera with the strains of the serovar Pomona, a complete absorption of all antibodies (anti-Pomona and anti-Mozdok) was achieved in seven cases using the strain Simon, and in one case with the strain S-23, whereas after absorption using the Pomona strain, the residual antibodies were still present in all sera, and also in the majority of them when they were absorbed using the strains S-23 and Pöstényi. In this context, the Pomona infection was determined in the case of eight patients. Hence it follows that not all strains of the Pomona serovar were suitable for the AATs. CONCLUSION: The presence of the human Mozdok leptospirosis was confirmed for the first time by the use of the agglutinin absorption test. A clear correlation between the habitat areas of the A. agrarius and the patients who were infected with the strains of the Mozdok serovar was determined.

[Possibilities for laboratory diagnosis of leptospiroses]. / Moznosti laboratórnej diagnostiky leptospiróz.

Leptospiroses are worldwide spread zoonoses caused by hydrophilic bacteria of the genus Leptospira. Humans can be infected by contact with an infected animal or indirectly via staying in a contaminated environment (water, wet soil), in natural foci, while working outdoors, or while doing outdoor sport and leisure activities. Leptospirosis may manifest as a mild flu-like illness or in a severe febrile form (meningitis, pulmonary haemorrhage, hepato-renal syndrome, or myocarditis). Presented are the laboratory diagnostic methods for leptospiroses with their advantages and disadvantages. In practice, serological diagnosis by the microscopic agglutination test, which is also a confirmatory test, and, less often, the enzyme-linked immunosorbent assay are used. Methods of molecular biology are being introduced for direct rapid diagnosis of leptospiroses, in particular the polymerase chain reaction and its modifications, which allow the detection of Leptospira infection in acute phase.

DNA relatedness and serotyping of Leptospira strains.

BACKGROUND: In a previous study (5), the constructed phylogenetic tree for leptospires belonging to 12 different serovars common in Central Europe made the prediction of serovar from knowing the genotype and vice versa possible. OBJECTIVE: The study was aimed at investigation of the usefulness of such procedure to distinguish in between at present to us available and worldwide accepted reference strains of pathogenic Leptospira serovars. MATERIAL AND METHODS: One hundred and seventy seven Leptospira strains representing different serovars were tested. DNA fingerprints of these strains were performed, digitally captured and as described earlier of those phylogenetic tree using different fingerprinting software was constructed using UPGMA clustering method with band matching by the Dice coefficient (5). RESULTS: At this tree, 145 of 177 Leptospira strains tested each took a unique position, and the remaining 32 strains were distributed at 15 different positions (each of 14 positions taken by two different strains and one position taken by four strains). CONCLUSION: The constructed phylogenetic tree likely to be very useful in prediction of Leptospira serovar in most cases of an infection so the saving time and being helpful in serovar identification of the pathogenic agent (Fig. 1, Ref. 9).

Long-term trends in the epidemiology of human leptospirosis (Slovak Republic, 1954-2006).

The purpose of this study was the observation of eventual shifts in some basic epidemiological features of human leptospirosis in Slovakia over a long period of time. Epidemiological traits of human leptospirosis were evaluated in three decades: 1954-1963 (822 cases), 1976-1985 (477 cases) and 1997-2006 (310 cases). The evaluation encompassed the aetiologic structure of cases, the incidence rate of diseases, men-to-women ratio of patients, as well as the professional and seasonal distribution of patients. The most affected feature was the incidence rate, which decreased by more than 70% over time. Another considerable change was the proportion of different major serological types of leptospirosis. The initially prevailing Sejroe infections fell to 4th place in the percentage rank, while Grippotyphosa disease, which was ranked 2nd place at the beginning of the observations, became dominating. Icterohaemorrhagiae disease climbed from 4th place to 2nd place. Third place was reserved by the Pomona + Tarassovi infections during the whole study period. A distinct tendency of age- and gender-specific incidence rate compared to the proportion of leptospirosis by age and gender was noticed; only minor alterations of the values of both parameters were registered over time. The overall value of the men-to-women ratio (MWR) of diseased persons was virtually within the same range in all three time periods but varied according to different age groups. The MWRs relative to the causal Leptospira serovars were stable over time but markedly differed among distinct serovars. Incidence rates related to age and aetiology showed different trends for the major serological types of leptospirosis. Changes were observed in the professional distribution of leptospirosis: there was an important proportional decrease of cases among farmers and field workers, an increase among house-wives + pensioners, but only some slight changes in abattoir workers/butchers, pupils + students and workers. The seasonal distribution of patients did not show any remarkable changes; the maximum percentage of cases occurred during the period extending from July to November during the whole period of observation. The epidemiological features of human leptospirosis underwent important shifts in the Slovak Republic over a 50-year period of time. They were very closely related to economical, social and political changes, which are discussed in this paper. The results may be useful for specialists in other European countries.

[Results of surveillance of human leptospirosis in Slovakia 1986-2005]. / Vý sledky surveillance humánnych leptospiróz, Slovensko 1986-2005.

STUDY OBJECTIVE: Presentation of results of surveillance of human leptospirosis over the last 20 years. MATERIAL: The data sources were the annually published results of leptospirosis surveillance and study of natural focuses of these infections. RESULTS: We describe trends in the overall incidence of leptospirosis, distribution of cases by etiological agent and type of leptospirosis, morbidity rate, distribution of patients with different types of leptospirosis by age and occupation/socio-economic group, leptospirosis seasonality and clinical forms. Some epidemiological measures that were taken are specified. Intensity of the epizootic process of leptospirosis in natural foci is correlated with the incidence of the disease in humans. Basic data from surveillance of leptospiral infections in farm animals as the source of infection for humans are presented. CONCLUSION: The significance and usefulness of conducting regular leptospirosis surveillance are emphasized.

Utility of phylogenetic studies in the identification of Leptospira strains.

A phylogenetic tree, which distinguishes between the serovars and serogroups of leptospires common in Central Europe was constructed using an established RAPD procedure together with digital reading and evaluation (using different computer software programs) of the generated amplified DNA patterns. The application of this procedure has revealed a consistent correspondence between serogroup and genotype (position in constructed tree) in 69 cases, and serovar and genotype in 72 cases, of wild strains of leptospires. There was an agreement between serovar and genotype in cases of strains of Grippotyphosa, Pomona, Mozdok, Arborea and Sorexjalna as well as between serogroup and genotype in cases of Australis, Bataviae and Sejroe. With the procedure used in this study, it was not possible to distinguish between reference strains of serovars Jalna, Bratislava and Lora (all serogroup Australis) as well as between serovars Icterohaemorrhagiae and Copenhageni (both of serogroup Icterohaemorrhagiae). In contrast to this, wild strains belonging to serogroup Sejroe were distributed between Polonica, Istrica, Saxkoebing and Sejroe serovars. Endemic strains of leptospires tested, were also distinguishable.

[Specificity of serologic diagnostic tests for legionellosis]. / Problém specificity sérologických diagnostických testov pri legionelóze.

Immune rabbit sera and sera of patients with legionellosis, leptospirosis, Q fever and antibodies against Borrelia burgdorferi were examined in cross reactions. In currently used serological tests, indirect immunofluorescence and the microagglutination test no antibodies were detected which could cause difficulties in the interpretation of results in the diagnosis of legionellosis. In one patient a mixed infection with Q fever and legionellosis was found.

[Leptospirosis in dogs in Slovakia]. / Premorenie psov na Slovensku leptospírami.

Out of 571 serologically investigated dogs from Bratislava and all regions of Slovakia and Moravia, 26.1% presented antibodies reacting with leptospiral antigens. The lowing seropositivity in dogs from Bratislava was found in lap dogs--11.1%, the highest one in farm dogs--35.3% (Tab. I). The seropositivity of police dogs from different regions of Slovakia and Moravia varied from 24.0% to 47.6% (Tab. II). The age of dogs progressing, their seropositivity rose from 3.1% in animals younger than one year to 33.3% in those aged seven years and in older ones (Tab. III). The predominant serovars causing infections of dogs were L. grippotyphosa (58.9% of infections) and serovars of both Sejroe (21.0%) and Icterohaemorrhagiae groups (15.3%). The serovars bratislava/jalna, pomond/mozdok and tarassovi were only rarely involved in these infections, canicola and arborea only exceptionally (Tab. IV). At present, in the Czechoslovak conditions the dogs do not play a significant role as reservoirs of human leptospirosis, and the predominant serovars may only cause irregular and short-time lasting leptospiruria. This is why the reasonableness of actual vaccination of dogs, possibly with the exception of the vaccination against Weil's disease, is doubtful. Dog is a good indicator of the distribution of different leptospiral serovars in its environment. It seems reasonable to practice occasionally serological investigations of these animals in order to detect possible changes in infecting leptospiral serovars. Consequently, adequate epizootological measures could be taken (e. g. modification of the composition of a leptospiral vaccine for dogs).

[Effort to eradicate a focus of tularemia and leptospirosis]. / Versuch der Tilgung eines Naturherdes der Tularämie und der Leptospirose.

An endemic area of tularemia and field fever was submitted to a complex surveillance over a period of more than 10 years. In 1977 an epizootic outbreak of tularemia appeared in all four localities studied. The infection rate in small mammals exceeded 10%. F. tularensis was isolated also from different species of ectoparasites. During 1978 the epizootic stopped in 3 foci, but continued to spread in one. Rodenticides (zinc phosphide) were therefore applied in this focus twice annually for 2 1/2 years. Since 1978, tularemia infections disappeared in small mammals as well as in man. The application of rodenticides did not influence the circulation of Leptospira grippotyphosa in small mammals. Their infection rate ranged between 3 and 6%. The reason for this striking difference is seen in the different pathogenesis and transmission of both infections.