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PURPOSE: To report retrobulbar neuritis caused by Borrelia afzelii culturally proved from cerebrospinal fluid (CSF). METHODS: A 23 year old female underwent ophthalmologic, laboratory and other auxilliary examinations. RESULTS: CSF cultures grew spirochetal microorganisms, serotyped by monoclonal antibodies as Borrelia afzelii. Following the serological and cultural results, treatment with doxycycline 200 mg daily was started and kept for three weeks. Gradual improvement of the visual acuity of the right eye was observed with full recovery to 20/20. CONCLUSIONS: Borrelia infection should be considered in the differential diagnosis of retrobulbar neuritis. CSF should be examined also culturally. (Ref. 5.)

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The diagnosis of Lyme borreliosis, except in cases characterised by pathognomonic clinical manifestation, usually requires confirmation by means of microbiological diagnostic assay, mainly by antibody detection methods. In our study antibodies to B. burgdorferi were tested in neurological patients with suspected Lyme borreliosis, depending on syndrome and clinical diagnosis. Antibodies were tested with IFT, ELISA and immunoblot. Blood samples of patients tested with IFT and ELISA tests were positive in 88 patients. Positive indirect immunofluorescence tests were found in 83 patients; in 5 patients the antibody level was borderline. Of these, 40 were positive also in ELISA but a correlation between IF titers and ELISA-positivity was not established. The immunoblot method confirmed specific antibody positivity in 36 of 88 patients (45.45%) who were positive (or borderline positive) in the indirect IF test, and in 28 of 40 (70%) ELISA-positive patients. Antibody specificity was found in 8 indirect IF-positive patients who were ELISA negative. This may be explained by the higher immunoblot sensitivity in comparison with ELISA. The Lyme borreliosis diagnosis was clinically established in 19 patients; antibodies to B. burgdorferi were only found in 13 patients in all three tests, and in 4 patients only in the indirect IF test. The results of serological tests for antibodies to B. burgdorferi should be interpreted with caution, as the tests are not standardized and may show false positive or false negative results. A two-step serological examination with the immunoblot test is recommended, whereby some nonspecific reactions may be eliminated. The results of serological tests have only supportive value and cannot be deemed conclusive when establishing an etiological diagnosis.

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At the beginning of new millennium, Lyme borreliosis is still the subject of intensive research, polemic discussions and open questions. The authors present minute analysis of the issues associated with Lyme borreliosis, concentrating on biological aspects and taxonomic classification of the agent, co-transmission and co-infection, diagnostic criteria and their validity, laboratory diagnostics an therapy of the disease including perspectives of active immunisation in the future. Special attention is paid to open questions in clinical and laboratory diagnostics of the disease and to the prospects for the near future. The authors also discuss the importance of international and interdisciplinary cooperation in the process of articulation of diagnostic criteria and recommended procedures for laboratory diagnostics. (Ref. 17.)

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UNLABELLED: Antibody response in infections with Borrelia burgdorferi sensu lato is generally considered to be slow and delayed, but exact studies concerning this question are hardly found in the scientific literature. During 1994-1998 at least two serum samples were submitted for serological testing from more than 1200 patients. An immunofluorescence test was performed paralelly with two pools of antigen (B. bg.s.s. + B. afzelii, and two serological different strains of B. garinii, all of local origin). In 92-96% of patients no change of antibody level was found in repeated tests, about 20% of them being negative (< 1:512). In 2-4% of cases a significant increase and in less than 1% a decrease or a second wave of the rise of the antibody level was observed. From the 58 patients with increasing antibody levels 9 had ECM (mostly children), 2 cardiovascular, 8 musculo-skeletal and 12 neurological disorders. In the remaining cases the diagnosis was not specified (A 69). Only in 9 cases a rise of the titer appeared during 3 weeks after the first negative sample, at contrary in 7 cases no rise of the titer was seen in that time. 2 patients were still after 1 month, 3 after 3 months and 1 even after 7 months (patient with a positive CSF culture) serologically negative. A 2 titer step decrease was observed in 4 cases during 1 year and once a 3 titer decrease after 4 years. CONCLUSION: A rise of antibodies can be seen in all clinical manifestations of Lyme disease with prevalence in the early types of the disease. The rise of antibodies is often delayed or even inhibited, apparently mostly by an early efficient treatment. Whether a late rise of antibodies or its long persistence indicates a latency of the infection, requires more detailed studies. (Tab. 5, Ref. 6.)

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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.

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The first four strains of Borrelia burgdorferi isolated in Slovakia from ticks and mice were studied using monoclonal antibodies, the polymerase chain reaction (PCR) with 16S rRNA specific primers and plasmid profiles. Two tick isolates were typed as Borrelia garinii, one strain isolated from Apodemus flavicollis was found to be B. afzelii and the fourth tick isolate reacted as a mixed culture of B. garinii and B. afzelii. All four strains harboured several plasmids ranging from 6-50 kbp including a plasmid with a size of approximately 41 kbp.

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We have developed a one-point microcapsule agglutination test (MCAT) for the serodiagnosis of leptospirosis. The MCAT kit was evaluated for use in humans by six WHO Collaborating Centres for Reference and Research on Leptospirosis. The laboratories classified their serum samples on the basis of the microscopic agglutination test (MAT) and the following screening tests: enzyme-linked immunosorbent assay (ELISA), macroscopic (slide) agglutination test, or the complement fixation test. The MCAT may in some instances give a positive result earlier in the course of the disease than MAT or the ELISA IgM; on the other hand, it did not detect antibodies against some serovars, for example, those of the Sejroe or Australis serogroup in Slovakia. In contrast, however, the MCAT detected antibodies to serovar hardjo (the same serogroup as Sejroe) in patients from the United Kingdom and the Russian Federation.

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The ecological index (EI) as defined by Schulze et al. was determined in three localities of western Slovakia (Malacky, Podunajské Biskupice, Olichov) using the following criteria: suitability of the habitats for the occurrence of Ixodes ricinus ticks and their number, accessibility of habitat for men (I. phase); furthermore the abundance and infection rate of ticks (II. phase). A scoring procedure (point values of 1-5 for each parameter) was used to evaluate the EI of three localities studied. Based on obtained values it is suggested to differentiate foci as those with a definite (Malacky), potential (Podunajské Biskupice, Olichov), minimal and unprobable risk of transmission. The EI is considered a helpful method for assessing areas of risk for the transmission of Lyme borreliosis, indicating also the necessity of appropriate preventive measures.

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The authors present examples demonstrating that in the last decades we witnessed the discovery of new nosological units and their infectious agents, the detection of the infectious origin of "old" diseases the aetiology of which was still recently obscure, infectious attacks of people by diseases which were considered animal infectious only, as well as the more frequent incidence of opportune infections due to impairment of the immune state of the human organism. These changes have many reasons such as the development of properties of microorganisms, lifestyle changes of the human population, advances in medicine, and last but not least improving microbiological diagnosis. Attention is drawn to the fact that even in a time when so-called diseases of civilization are in the foreground of interest our awareness of infectious diseases must not slacken. Only all-round international surveillance can prove successful.

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Serological examination of 420 domestic animals for the presence of antilegionella antibodies indicates their high exposure to legionellae. On examination by the microagglutination reaction with a serum dilution of 1:64 or more the highest positive values were recorded in horses which reacted with antigens of L. pneumophila 1-14 in 36.2% and with antigens of another 19 types of legionellae in 47.8%. In pigs positive values recorded in 16.2% and in 21.1%; in cattle in 3.8% and 29.5%, in sheep in 7.5% and 11.3% and laboratory rabbits were quite negative. The importance of these findings with regard to the possible role of animals in the ecology of legionellae is obscure.

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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).

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Five leptospiral strains were isolated from bovine kidneys during a cultural survey for pathogenic leptospires in Nigeria. Preliminary test results indicated that the five strains were identical and serologically heterologous to the other members of the Pyrogenes serogroup. Further examination of the strains by the cross-agglutinin absorption test, factor analysis and restriction endonuclease analysis confirmed that the strains constitute a new serovar. It is therefore proposed that this strain be recognised and designated as serovar nigeria, type strain Vom.

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Examination of ticks for the presence of F. tularensis resulted in the isolation of 29 strains, predominantly from the species D. reticulatus (19x), further from I. ricinus (8x) and from H. concinna (2x). The results confirmed the activity of the known endemic focus in the area of Podunajské Biskupice and revealed three new foci in the localities Smolenice (district of Trnava), Olichov (district of Nitra), and Plást'ovce (district of Levice). In the focus of Podunajské Biskupice the mean infestation of ticks of the species D. reticulatus was around 1.4% over the years of investigation. With regard to the one-year developmental cycle, D. reticulatus can be considered an appropriate indicator of current epizootic activity of the focus, while the occurrence of I. ricinus is rather an indicator of the geographic spread of foci. The third part of the study addresses questions of the ecology of ticks. It analyzes their biological characteristics in our conditions, their spread and bionomy, particularly from aspects related with their epidemiological significance as vectors of Lyma borreliosis and tularemia. Systematic follow up of infestation of ticks with causative agents of borreliosis and tularemia in established endemic foci as well as in epidemiologically indicated localities is an inevitable part of modern surveillance of these infections.

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Cultures of B. burgdorferi, their supernatants as well as washed cells revealed in vitro a considerable antitumor activity against cells of Gardner lymphoma. Németh-Kellner lymphoma and LP-2 plasmacytoma. In in vivo tests an inhibition of tumor growth was evident, even if the treatment was started on day 4 after implantation of the tumor. The best results were obtained with the supernatants of the cultures.

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Among 30 strains of leptospires isolated from samples of sewage taken before and during treatment at two sewage plants in England, only one appeared to belong to Leptospira interrogans, the species that comprises the leptospires that are pathogenic to man and animals. That strain, Compton 746, was isolated from settled sewage, before treatment at a treatment plant that deals mainly with human sewage. It was shown serologically to belong to serogroup Tarassovi and appears to represent a new serovar that has been named mogdeni after the name of the sewage plant, Mogden, from which it was isolated.

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