Anaplasma phagocytophilum is associated with diseases of goats, sheep, cattle, dogs and horses. In the beginning of the 1990s it was identified as a human pathogen, causing human granulocytic anaplasmosis (HGA) in the USA, Europe and the far east of Russia. A. phagocytophilum is maintained in nature in an enzootic cycle including ticks as the main vector and a wide range of mammalian species as reservoirs. Ixodes ricinus and I. persulcatus ticks were collected in Estonia, Belarus and the European part of Russia and screened for the presence of A. phagocytophilum by real-time PCR. Positive samples were found only among I. ricinus, in 13.4% in the European part of Russia, 4.2% in Belarus, 1.7% in mainland Estonia and 2.6% on Saaremaa Island. Positive samples were sequenced for partial 16S rRNA, groESL and ankA genes and phylogenetic analyses were performed. The results showed that A. phagocytophilum circulating in Eastern Europe belongs to different groESL lineages and 16S rRNA gene variants and also consists of variable numbers of repetitive elements within the ankA gene.

The presence of Babesia spp. was studied in 2603 Ixodes ricinus and Ixodes persulcatus ticks collected at seven sites in Estonia. By reverse line blot screening, Babesia spp. was detected in 36 (1.4%) ticks, among them 18 (0.7%) were further recognized by a Babesia microti probe, 3 (0.1%) by a Babesia divergens probe, and the other 15 (0.6%) were recognized only by the universal Babesia spp. "catch all" probe. Sequence analyses of 6 of these 15 samples revealed that all of them belonged to Babesia sp. EU1. B. microti was detected in both tick species I. ricinus and I. persulcatus at the seven sites, whereas B. divergens-like and Babesia sp. EU1 were found only in I. persulcatus and I. ricinus, respectively. Genetic characterization based on partial 18S rRNA showed that the Estonian sequences of B. microti, B. divergens-like, and Babesia sp. EU1 share a high rate of similarity and are closely related to sequences from other European countries, Siberia, and United States. The present study demonstrated for the first time the existence and distribution of Babesia spp. in I. persulcatus and I. ricinus ticks in Estonia.

BackgroundA single-tier immunoassay using the C6 peptide of VlsE (C6) from Borrelia burgdorferi sensu stricto (Bb) has been proposed as a potential alternative to conventional two-tier testing for the serologic diagnosis of Lyme disease in the United States and Europe.ObjectiveTo evaluate the performance of C6 peptide based multiplex Phosphorescence Analysis (PHOSPHAN) for the serologic confirmation of Lyme borreliosis (LB) in Russian patients.MethodsSerum samples (n = 351) were collected from 146 patients with erythema migrans (EM); samples from 131 of these patients were taken several times prior to treatment and at different stages of recovery. The control group consisted of 197 healthy blood donors and 31 patients with other diseases, all from the same highly endemic region of Russia. All samples were analyzed by PHOSPHAN for IgM and IgG to Bb C6, recombinant OspC and VlsE proteins, and C6 peptides from B. garinii and B. afzelii.ResultsIgM and IgG to Bb C6 were identified in 43 and 95 out of 131 patients (32.8 and 72.5%, respectively); seroconversion of IgM antibodies was observed in about half of the patients (51.2%), and of IgG antibodies, in almost all of them (88.4%). Additional detection of OspC-IgM and VlsE-IgM or IgG to C6 from B. garinii or B. afzelii did not contribute significantly to the overall sensitivity of the multiplex immunoassay.ConclusionsThe multiplex phosphorescence immunoassay is a promising method for simultaneously revealing the spectrum of antibodies to several Borrelia antigens. Detection of IgM and IgG to Bb C6 in the sera of EM patients provides effective serologic confirmation of LB and, with high probability, indicates an active infection process.

The etiological structure of infections transmitted by Ixodes ticks (ITIT) was established in 467 (out of 522) patients of the Perm Regional Hospital for Infectious Diseases with the help of Laboratory Techniques Set, which included ELISA, PCR and PHOSPHAN М. There was stated that ITIT occurs most frequently - 235 (45, 0%) patients, tick borne encephalitis (TBE) -more rarely: 54 (10.4%), human granulocytic anaplasmosis (HGA) - 26 (5.0%) and human monocytic ehrlichiosis (HME) - 9 (1.7%). Various mixt-infections were detected in 143 patients (27.4%). In 55 patients (10.5%) the etiology of the disease remained to be unexplained. The clinical course of ITIT caused by genospecies B. afzelii and B. garinii is described.

In this report, we evaluated the performance of C6 peptide based multiplex Phosphorescence Analysis (PHOSPHANTM) and Polymerase Chain Reaction (nested PCR) for laboratory diagnosis of Ixodid Tick-borne Borrelioses (ITBB). The study was conducted on 155 patients with localized and disseminated stages of the disease, the cases of mixed infection with ITBB and human granulocytic anaplasmosis including. Positive PHOSPHAN reactions were observed in 78 ± 7.7% of patients with erythema migrans (EM) and 91 ± 11.7% of patients without cutaneous manifestations of the disease. The frequency of PCR positive samples was lower, 26 ± 8.2% and 72 ± 17.1% respectively. The maximum frequency of positive samples detected by both methods was mainly observed at 2 - 4 week from the onset of the disease (or 22 - 35 day after tick bite). In general, PHOSPHAN provided serologic confirmation of the disease in 52 of 55 (94.5 ± 6.2%) patients, whose blood contained Borrelia DNA. Only 3 patients tested positive in PCR (1 - with EM and 2 - without this skin manifestation) were seronegative. These data confirmed the high efficiency of PHOSPHAN method for serologic verification of ITBB both at localized and disseminated stages of the disease. The use of PCR (in addition to PHOSPHAN) is appropriate within a certain period of time (no later than 2 - 3 weeks from the onset of the disease) to clarify the diagnosis in seronegative patients having clinical signs of disseminated non-cutaneous form of ITBB, or atypical cutaneous manifestations of erythematous form of the disease.