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The aim of the present study was to determine the frequency and spatial distribution of the Borreliaspecies in wild rodents in the Czech Republic. In total, 293 muscle tissue samples and 106 sera from 293 wild rodents captured in North Bohemia and North-East and South Moravia were examined for the presence of Borreliaspp. and antibodies. Muscle samples were investigated with real-time polymerase chain reaction (PCR) with a recAprimer set, with DNA quantification and melting curve analysis, and with restriction fragment length polymorphism (RFLP) analysis of the 5S–23S rDNA intergenic spacer. Infection with Borrelia burgdorferisensu lato was found in 16.4 of the muscle samples. The most abundant genospecies was Borrelia afzelii(11.3), followed by Borrelia burgdorferisensu stricto (4.8) and Borrelia garinii(0.7). Borreliainfection was more frequently observed in Clethrionomys glareolusthan in Apodemusspp. Sera were analyzed with an enzyme-linked immunosorbent assay (ELISA) test, yielding the total seropositivity rates of 24.5 for anti-BorreliaIgM antibodies and 25.5 for IgG antibodies. Total seroprevalence was higher in Apodemusspp. than in C. glareolus. In conclusion, our data indicate that in the Czech Republic small wild rodents can serve as hosts for B. burgdorferis. s. as well as for B. afzelii. [ABSTRACT FROM AUTHOR]

Objectives: For the proper diagnosis of toxoplasmosis it is essential to determine the stage of the infection, for which the most preferred method is IgG avidity test. The avidity index (AI) should initially be low (AI≤0.3) in the acute phase and increase during the infection. However, persistent low avidity can occur in patients with latent toxoplasmosis, which can complicate the interpretation of the results. The aim of the study is to explain the causes of this phenomenon., Methodology: A retrospective study was carried out with 717 serum samples collected from 442 patients from the categories of pregnant and non-pregnant women, men, and newborns + infants (age < 0.5 year). The trends of AI kinetics were evaluated in repeatedly examined patients. The frequency of cases with low avidity was compared in individual categories of patients and in groups of people with acute and non-acute toxoplasmosis., Results: The proportion of patients with initially low avidity was 42.1% in the acute toxoplasmosis group while it was 13.0% in the non-acute groups. In uninfected newborns with anti-Toxoplasma antibodies transmitted from the mother, a decrease in IgG avidity levels over time was observed, resulting in 29.2% of samples showing low (improper) avidity. While the dynamics of IgG avidity and the frequency of cases of improperly low avidity were similar in men and pregnant and non-pregnant women, the category of newborns and infants differed substantially for these indicators., Conclusions: Due to acceptable specificity and negative predictive value, high avidity can rule out acute toxoplasmosis, but moderate sensitivity complicates the possibility of its confirmation. The results of the avidity test must be interpreted in the context of the results of other methods., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Kodym et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The study compares diagnostic parameters of different commercial serological kits based on three different antigen types and correlates test results with the status of the patient's Borrelia infection. In total, 8 IgM and 8 IgG kits were tested, as follows: enzyme-linked immunosorbent assay (ELISA) (Euroimmun) based on whole-cell antigen, 3 species-specific enzyme immunoassays (EIAs) (TestLine), Liaison chemiluminescence (DiaSorin), ELISA-Viditest (Vidia), EIA, and Blot-Line (TestLine) using recombinant antigens. All tests were performed on a panel of 90 samples from patients with clinically characterized borreliosis (53 with neuroborreliosis, 32 with erythema migrans, and 5 with arthritis) plus 70 controls from blood donors and syphilis patients. ELISA based on whole-cell antigens has superior sensitivity and superior negative predictive value and serves as an excellent screening test, although its specificity and positive predictive values are low. Species-specific tests have volatile parameters. Their low sensitivity and low negative predictive value handicap them in routine diagnostics. Tests with recombinant antigens are characterized by high specificity and high positive predictive value and have a wide range of use in diagnostic practice. Diagnostic parameters of individual tests depend on the composition of the sample panel. Only a small proportion of contradictory samples giving both negative and positive results is responsible for discrepancies between test results. Correlation of test results with the patient's clinical state is limited, especially in the erythema migrans group with high proportions of negative and contradictory results. In contrast, IgG test results in the neuroborreliosis group, which are more concordant, show acceptable agreement with Borrelia status., (Copyright © 2018 Kodym et al.)

Sera collected from healthy individuals from the general population in the Czech Republic during repeated cross-sectional surveys were analyzed. Samples collected in the same six districts in two time periods, 1978-1989 and 2001, were compared. The study subjects were divided into six age categories between 10 and 59 years. Overall, 434 samples from 1978-1989 and 270 samples from 2001 were screened for Anaplasma phagocytophilum (AP) and Borrelia burgdorferi sensu lato (BB). The anti-AP positivity rates were 13.1% and 11.5% in the first and second period, respectively, and did not differ significantly between the periods (P = 0.559). The anti-BB antibodies were detected in 33.9% and 14.8% of study subjects, respectively. The positivity rates were significantly lower in the second period (P<0.001). No considerable changes were observed in the sex distribution of positive findings between the two periods. The highest positivity rates of anti-AP antibodies were found in the 10-14 year age group: 16.0% in 1978-1989 and 16.7% in 2001. The age distribution of the anti-AP antibody positivity rates did not change substantially (P = 0.872). In 1978-1989, the lowest anti-BB antibody positivity rate (26.7%) was found in the 10-14 year age group, with a gradual increase with age to 41.1% in 50-59 year-olds. In 2001, the positivity rate in the 10-14 year age group was 26.2% and was not significantly different from that in the first period (P = 0.955). However, the positivity rates in the older age groups 15-59 years decreased significantly (P<0.001) and varied between 8.3% and 15.1%.

The aim of our study was to find out the optimal conditions for short-term storage of cerebrospinal fluid (CSF) samples for direct diagnosis of Lyme disease. A mixture of Borrelia-negative CSFs spiked with a defined amount of cultured Borrelia garinii was used. Borrelia stability was investigated over 7 days at four different temperatures [room temperature (RT), +4, -20 and -70 °C]. Quantitative changes in CSF Borrelia were measured by quantitative PCR (qPCR), and morphological changes in the spirochetes were observed by transmission electron microscopy (TEM). These qPCR results were statistically evaluated. We found +4 °C to be an optimal temperature for short-term storage of CSF samples intended for TEM observation. There was no significant difference between the temperatures tested in the average quantity of Borrelia measured by qPCR. On the contrary, electron optical diagnosis of frozen samples and samples stored at RT showed destructive morphological changes and decreased spirochete counts. Our results show that optimal conditions for the pre-analytical phase of investigation of one type of material can differ depending on the diagnostic method employed.

The subject of this study is serological screening of blood and CSF (cerebrospinal fluid) samples for the presence of borrelial and ehrlichial antibodies. A total of 165 patients suspected to be at risk of Lyme disease were tested. Indirect immunofluorescence and enzyme immunoassay were used as diagnostic methods. Ehrlichial antibodies were detected in 36 (21.8%) patients. Borrelial antibodies were found in 70 samples (42.4%). The widening range of tick-borne diseases brings about the need for more data on these zoonoses.

The aim of this study is to present molecular, serologic, and clinical findings for dogs that were naturally infected with Anaplasma phagocytophilum or Borrelia burgdorferi sensu lato (s. l.) in the Czech Republic. This data can provide information relevant to human infection. In total, blood samples from 296 dogs and 118 engorged ticks were examined. Samples were tested for A. phagocytophilum using polymerase chain reaction (PCR) amplification, nested PCR, and direct sequencing of the 16S rDNA, and for B. burgdorferi s. l. using PCR amplification of the 16S rDNA and restriction fragment length polymorphism analysis of the 5S-23S rDNA intergenic spacer. In addition, blood samples were screened for antibodies to these bacteria. Ten (3.4%) dogs were PCR-positive for A. phagocytophilum. Morulae of A. phagocytophilum in granulocytes were found in two of these dogs. Nine of the PCR-positive dogs had clinical signs related to anaplasmosis. Statistically significant differences in the PCR detection rates were found between breeds and between symptomatic and asymptomatic dogs. Infection with Borrelia garinii was detected by PCR in a dog with meningoencephalitis. DNA of A. phagocytophilum and B. burgdorferi s. l. (B. garinii or Borrelia afzelii) was detected in 8.5% and 6.8% of ticks, respectively. Immunoglobulin (Ig) G seropositivity to A. phagocytophilum was 26%. Significant differences were found with respect to breed and gender. IgM and IgG antibodies to B. burgdorferi s. l. were detected in 2.4% and 10.3% of dogs, respectively. Our findings suggest that the exposure to B. burgdorferi s. l. exists in dogs in the Czech Republic, and exposure to A. phagocytophilum is common.

Anaplasma phagocytophilum has been first isolated from the blood of two Czech patients simultaneously with a cultivation of Borrelia burgdorferi sensu lato from their erythema migrans lesions. Cultivation of different Borrelia spp. from 12 erythema migrans biopsies, from 2 blood, one liquor and one placenta sample in BSK-H medium was successful. Adapted conventional methods targeting 16S rRNA and OspA genes for real-time polymerase chain reaction (PCR) and partial sequencing of these genes together with microscopical examinations of the blood smears provided a direct detection of the B. afzelii, B. burgdorferi, B. garinii, B. valaisiana and B. bissettii in the skin, B. garinii in the blood, placenta and liquor in 24 (36.3 %) patients, and A. phagocytophilum in 10 (15 %) patients with erythema migrans. Positive indirect IgM immunofluorescence against Anaplasma sp. was obtained in 7 cases, specific IgG antibodies were detected in 12 patients. Three women suffering from erythema migrans in the first trimester had positive PCR for Anaplasma and/or for Borrelia in the blood and two of them, later, in the placenta. Interpretation of laboratory data can bring important contribution to establishing the role of Anaplasma sp. in erythema migrans and forming the principle of precaution with laboratory diagnosis during pregnancy which always should be reflected in the resistance of Anaplasma sp. toward penicillins.

The aim of the present study was to determine the frequency and spatial distribution of the Borrelia species in wild rodents in the Czech Republic. In total, 293 muscle tissue samples and 106 sera from 293 wild rodents captured in North Bohemia and North-East and South Moravia were examined for the presence of Borrelia spp. and antibodies. Muscle samples were investigated with real-time polymerase chain reaction (PCR) with a recA primer set, with DNA quantification and melting curve analysis, and with restriction fragment length polymorphism (RFLP) analysis of the 5S-23S rDNA intergenic spacer. Infection with Borrelia burgdorferi sensu lato was found in 16.4% of the muscle samples. The most abundant genospecies was Borrelia afzelii (11.3%), followed by Borrelia burgdorferi sensu stricto (4.8%) and Borrelia garinii (0.7%). Borrelia infection was more frequently observed in Clethrionomys glareolus than in Apodemus spp. Sera were analyzed with an enzyme-linked immunosorbent assay (ELISA) test, yielding the total seropositivity rates of 24.5% for anti-Borrelia IgM antibodies and 25.5% for IgG antibodies. Total seroprevalence was higher in Apodemus spp. than in C. glareolus. In conclusion, our data indicate that in the Czech Republic small wild rodents can serve as hosts for B. burgdorferi s. s. as well as for B. afzelii.

The spread of borreliosis depends on geographical, environmental and climatic factors as well as on the pathogenesis of the causative agent of the group of Borrelia burgdorferi sensu lato. The rise in the incidence of the disease and emergence of new symptoms are of concern. Relationships between genospecies and symptoms, their geographical spread and possible interference of other pathogens are the subject of the present study. Eighty-seven patients with borreliosis from Central and Eastern Bohemia and Moravia were enrolled in the study. Forty-nine patients of group 1 showed clinical positivity, 21 patients of group 2 tested positive at PCR screening and 17 patients of group 3 were culture positive. Forty-eight patients and 17 isolated strains showed positivity for plasmids and the Borrelia burgdorferi sensu lato genome in conventional nested PCR. Borrelial genotypes and subtypes were detected by direct sequencing of OspA and OspC products. Quantitative data were determined from specific product melting temperature curves for real time PCR. Based on sequencing of the OspA gene, B. garinii (subtypes 6, 5, 4 and 3), B. burgdorferi s.s. and B. afzelii were detected in 14 (51.8%), 8 (29.6%) and 5 (18.5%) out of 27 Central Bohemian patients, respectively. Eastern Bohemian patients showed predominance of B. garinii subtype 5 and co-infection with Anaplasmaphagocytophilum in 7.6%. The predominant causative agent in 25 Moravian patients was B. afzelii (11 patients, i.e. 44%), followed by B. burgdorferi s.s. (9 patients, 36%) and B. garinii 5 patients, i.e. 20%). Sequences of two hypervariable regions of the OspA and OspC genes and distances in phylogenetic trees showed differences not only between genospecies and subtypes but also between wild strains detected by direct sequencing from patient specimens and in vitro cultured strains. The greatest differences were found for patients with long-term borrelial infection.

In the 2000 and 2001 years, there was performed a study which has given some insight on the knowledge of youths about ticks and tick-borne diseases. Two thousand seven hundred and sixty-three respondents from 6 to 26 years of age took part in this investigation. More than 98% of respondents knew about the existence of ticks. Almost 93% of children and 97% of adolescents reported that ticks feed on blood. Although the majority of respondents aged 10 to 26 years is convinced that ticks live on vegetation, 23% of them supposed that ticks jump on humans from trees. As many as 93.5% of youths knew that Lyme disease is transmitted by ticks. The main sources of information about Lyme disease for students and pupils older than 10 years of age are television and radio (40.9%) and the press (37.5%). The frequency of contact of young people with ticks is high--90.4% of children younger than 12 years, and 93.7% of youths from 10 to 26 years of age had at least once an attached tick. 56.1% of youths older than 10 years use oil to remove an attached tick. Almost 23.7% remove ticks with bare hands. People used those methods of tick removal although even children younger than 12 years of age knew that it was an incorrect habit. After removing, a tick is most frequently burned.