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Isolation of lysozyme from hemolymph of Alveonasus lahorensis (Acari: Parasitiformes, Argasidae) and Hyalomma marginatum (Acari: Parasitiformes, Ixodidae) with using ultrasound is described. It was shown that the bactericidal effect of the ultrasound-extracted lysozyme against Staphylococcus aureus and Micrococcus luteus significantly exceeded that of the chicken egg lysozyme and lysozyme from ticks without ultrasound exposure. Disintegration of the hemolymph cells increased lysozyme production.

Serological verification of ixodid tick-borne borrelioses (TBB) is carried out in Russia by using IFA with the standard antigen prepared from the strain Ip-21 B. afzelii by the Laboratory of Infection Vectors, Gamaleya Research Institute of Epidemiology and Microbiology, Russian Academy of Medical Sciences. This study is under way at approximately 50 research and practical institutions that are situated in 30 large administrative territories of the country. The nIFA data on 371 sera from 214 patients with TBB and 229 sera from controls who had other infections have been analyzed. Changes in the proportion antibody sera, mean geometrical and cut-off antibody titers in patients with TBB were compared with those in the controls. The values of seroconversion were analyzed in relation to the time lasted after the onset, the first serum collection, and an interval of reexamination. The specific results of tested of the coded sera were obtained during the international interlaboratory blind experiment. It is concluded that nIFA yields results that are suitable for TBB verification.

To investigate the relationships between tick-borne encephalitis (TBE) virus and the bacterial spirochaete Borrelia burgdorferi sensu lato in vectors with mixed infections, unfed adult Ixodes persulcatus ticks were collected by flagging from vegetation in southern-taiga forests of the Pre-Urals region of Russia where both infections circulate sympatrically. Prevalences of TBE and Borrelia infections in a total of 4234 ticks were compared over 5 years. No significant differences were revealed between the prevalence of Borrelia infection in ticks with and without TBE virus (29.4+/-7.8% vs 23+/-3.6%), or between the prevalence of TBE virus infection in ticks with and without Borrelia (24.0+/-6.6% vs 18.4+/-3.4%). In ticks with mixed infection (40/689 = 5.8%), concentrations of TBE virus and Borrelia were not significantly correlated with one another. Field observations showed parallel trends in the prevalence of these pathogens in tick populations from year to year (1993-1997) indicating that, in I. persulcatus with mixed infection, Borrelia and TBE virus do not seem to interfere with each other and are apparently not involved in any antagonistic relationships.

A special epidemiological experiment to prevent borreliosis in persons bitten by infected ticks was performed in 1992-1994 in the Russia's Perm region where Borrelia garinii and B.afzelii circulate, and Ixodes persulcatus tick is the sole vector transmitting these pathogens to human beings. Adult ticks were removed from the bodies of persons who had referred to health facilities for first aid. Vital preparations were made from the material obtained from the gut of each tick and examined microscopically (up to 250 microscopic fields per preparation). The patients bitten by infected ticks were divided into experimental and control groups and kept under special medical and serological control for 4-5 months. The patients of the experimental group received doxycycline (100 mg twice daily) for 3-5 days after ticks had bitten. Borreliosis was diagnosed by a combination of clinical and serological data. The control group consisted of 97 patients who took no antibiotics after ticks biting and 12 of them contracted borreliosis. In 823 cases Borrelia were not revealed while microscopically analyzing the ticks removed from the patients' bodies; in this group six patients contracted borreliosis. The morbidity rate (per 100 patients) in the experimental group was 1.1, i.e. 11 times lower than that in control group. Among the patients bitten by infected ticks and untreated with antibiotics, this index was 17.6 times higher than in the group bitten by ticks in which Borrelia were not found. There is no absolute probability of detecting the pathogen during a direct microscopic analysis of the preparation made from the tick removed from the body of a bitten patient. However, this rapid identification of Borrelia, followed by short-term antibiotic treatment for microbiological evidence is an effective tool for preventing patients from contracting borrelioses.

A total of 740 adult Ixodes persulcatus ticks were collected from the vegetation by flagging in Russian foci where Borrelia afzelii and B. garinii circulate, and 156 I. dammini ticks were collected in northwestern USA regions in foci with B. burgdorferi s.str. circulation. Smears prepared from the internal organs of ticks were stained according to Romanovsky-Giemsa and analyzed under a microscope at a x 1125 magnification. All borreliae in 250 microscopic fields were counted, and concentration of microbial bodies per 100 microscopic fields was determined. The general level of infection by Borrelia in both vectors was similar: 26.2 x 3.2 in I. persulcatus and 26.3 +/- 7 in I. dammini. However, the proportions of ticks with generalized infections differ considerably (12.9 +/- 4.8 in I. persulcatus compared with 2.4 +/- 4.8 in I. dammini; significance of difference t = 3.1). We did not reveal any definite increase in the proportion of ticks with borreliae in the salivary glands among ticks with high concentrations of microbial bodies in the gut. In 25 I. persulcatus ticks with generalized infections, series of actual numbers of borreliae (per 100 microscopic fields) found in the gut and salivary glands did not correlate with one another (r = -0.23). These results confirm our previous conclusion (Korenberg, 1994) that frequencies of generalized infection in main vectors of different ixodid tick-borne borrelioses are also different, which is probably due to peculiarities of relationships between spirochetes of each species and corresponding tick vectors. These factors can be responsible for differences in the ways of horizontal and vertical transmission of pathogens belonging to the group under study.

A direct microscopic analysis of fixed smears and live preparations was used to reveal whether spirochetes are present in the gut and salivary glands of adult Ixodes persulcatus ticks spontaneously infected with Borrelia garinii and B. afzelii. Unfed ticks collected from a vegetation, partially fed ticks removed from human bodies, and ticks deliberately fed on laboratory animals were studied. In each preparation, all spirochetes were counted in 250 microscopic fields, and their concentration per 100 microscopic fields was determined. A total of 1962 ticks were individually analysed. The methods used on the study allowed a reliable identification of Borrelia in the viscera of not only unfed, but also of partially fed ticks. The infection rate in ticks that started bloodsucking was slightly lower than in the unfed ticks. This was associated with the decreased spirochete concentration in the preparations made from the partially fed ticks. Borrelia were frequently found in the salivary glands of the unfed infected I. persulcatus. During the first two to three days of bloodsucking, neither the proportion of ticks with spirochetes in the salivary glands, nor the spirochete concentrations increase. Borrelia migration from the tick gut into the salivary glands during early bloodsucking is not a prerequisite for or even important for pathogen transmission with saliva. The transmission rate appears to depend on the baseline proportion of the unfed ticks carrying spirochetes in their salivary glands.(ABSTRACT TRUNCATED AT 250 WORDS)