Universiẗat, Diss., 2004--München.

Universiẗat, Diss., 2003--Jena.

Lyme borreliosis (LB) is caused by spirocheter of Borrelia burgdorferi sensu lato. There are different types of borrelia species and some differ in their ability to survive in the presence of the complement system. B. afzelii is complementresistant while B. garinii is complementsensitive. This is based on the ability to recruit immune regulators, such as factor H to the bacterial surface and prevent activation of the complement system. Some individuals may show anti-Borrelia antibodies without having developed clinical symptoms. This may indicate a more effective immune response against spirochetes. The aim of this study was to investigate differences in complement activation by measuring C3a and sC5b-9 with sandwich ELISA between two previously Borrelia-exposed groups; individuals with previous subclinical Lyme borreliosis (SB) and patients previously diagnosed with neuroborreliosis (NB), and a control group without signs of LB exposure. Samples analyzed in this study consisted of controls (Ctrl, n = 8st), SB (n = 60st) and NB (n = 22st). Plasma from the groups were activated with ACA1 and Lu59. To compare the relative increase between the groups, complement factor C3a and the soluble terminal complement complex, sC5b-9, were analyzed using sandwich-ELISA.The analysis of C3a and sC5b-9 showed higher activation with Lu59 than ACA1, which is consistent with previous studies. According to C3a-analysis, no significant differences were observed between the groups for neither ACA1 nor Lu59. According to sC5b-9-analysis, a significant difference between SB and Ctrl (p= 0,0081) for Lu59 was observed. Conclusion of the studie was that further studies are required to interpret how this complement activation affects LB from a clinical prespective.

In this study we examined the effect of the tick saliva from I. ricinus and the effect of recombinant protein IRS-2 from the saliva of I. ricinus on dendritic cells derived from the mice bone marrow. We studied their effect on the production of cytokines by dendritic cells after the stimulation by B. burgdorferi, their effect on the expression of genes, that participate in phagocytosis, and the impact of the tick saliva on phagocytosis of B. burgdorferi by dendritic cells.

During a period of 3 years, from March to July 2014, 2015 and 2016, ticks were collected from dogs and cats in shelter facilities for abandon animals in Česke Budejovice, South Bohemia. In total, 343 ticks were found on 106 pets: 67 domestic dogs and 39 cats. All collected ticks, that were identified as Ixodes ricinus and Ixodes hexagonus, were tested for the presence of spirochetes from Borrelia burgdorferi sensu lato (s.l.) complex, Rickettsia spp., Anaplasma spp, and Babesia spp using conventional PCR and nested PCR. Identification of pathogens was done by following sequencing of amplicons. Out of all tested ticks, 49,56% were proved to be infected at least with one pathogen. Co-infection of at least two different pathogens was determined in 18 ticks (5,2%). The aim of the present study was to estimate the role of accompanying animals (cats and dogs) in the circulation of ticks and tick-borne pathogens, to determine the frequency of pathogenic infections in dog and cat associated ticks, to evaluate the current risk of infection for dogs and cats, with respect torisk forhumans living in the area of České Budějovice.

Introduction: Inflammatory cardiomyopathy (DCMi) represents a non-familial form of dilated cardiomyopathy (DCM) and endomyocardial biopsy (EMB) is crucial for its diagnosis. Aims: To assess the prevalence of DCMi in patients with DCM of unclear origin, to evaluate the significance of serological tests for antibodies against infectious cardiotrophic agents and to analyze the effect of specific therapy guided by EMB results. Methods: EMB was performed in 56 subjects (mean age 52 ± 10 years) with DCM of unclear etiology and left ventricular (LV) ejection fraction (EF) < 40% with a history of heart failure less than 1 year. EMB samples were analyzed by immunohistochemistry, polymerase chain reaction (PCR) and electron microscopy. Results: Immunohistochemical examination revealed myocardial inflammation in 26 patients (46%), the PCR method detected genome of microbial agents in 32 patients (57%). Electron microscopy showed the presence of particles of microbial agents in 41 patients (73%). Serological blood tests found no IgM antibody positivity against any of the investigated microbial agents. Targeted antibiotic therapy in patients with evidence of Borrelia burgdorferi (Bb) genome in the EMB led to a reduction in LV size, improvement of LV EF and alleviate symptoms of heart failure. Conclusion: DCMi...

I describe procedures for the introduction of Borrelia burgdorferi, the spirochetal agent of Lyme disease, into larval and nymphal stage of the tick vector, Ixodes ricinus. The goal of this Mgr. Thesis is to find an optima system, that would reliably and reproducibly allow the infection of large numbers of ticks with the Lyme disease spirochete.

Interaction between mouse dendritic cells (DCs) and Borrelia afzelii spichochetes was studied on three different levels: phagocytosis of borrelia by DCs, production of cytokines by borrelia-activated DCs and the ablilty of DCs to activate CD4+ T cells. The effect of Ixodes ricinus saliva on each of these levels was examined. Tick saliva was shown to decrease the number of phagocosing DCs. The ability of borrelia-activated DCs to induce both proliferation and IL-2 production in specific CD4+ T cells was significantly reduced by tick saliva. And surprisingly, we have shown an inhibitory effect of I. ricinus saliva on the production of both Th1 (IL-6 and TNF-{$\alpha$}) and Th2 (IL-10) cytokines. Our data reveal a complex inhibitory effect of tick saliva on DC function.

The Lyme disease causing spirochaete Borrelia burgdorferi sensu lato is transmitted by ticks within the genus Ixodes. These ticks are liberal host seekers and parasitise mammals, birds and reptiles. Prior to this study, the distribution of I. ricinus ticks and Lyme disease was thought to be restricted to the southern half of Sweden. On the island Norrbyskär, located in the Bothnian Gulf, there were reports of a high incidence of tick infestation on humans. To investigate the occurrence of B. burgdorferi s.l. in these ticks and to characterise presumptive isolates at the molecular level we sampled a number of I. ricinus ticks. Three different isolates were obtained from two different ticks, NBS16 from a nymph and NBS23a and NBS23b from an adult female tick. The seabird associated tick I. uriae is circumpolar distributed in both hemispheres. On the island Bonden, which house one of the largest seabird colonies in the Baltic Sea, I. uriae were collected and surveyed for spirochaetes. One isolate of B. burgdorferi s.l. was obtained. This B. burgdorferi s.l. isolate is identical to the Lyme disease Borrelia strain NBS16 isolated from Norrbyskär. To investigate the role of seabirds in the epidemiology of B. burgdorferi s.l., I. uriae were collected from seabird colonies in the southern and northern hemispheres. Borrelia DNA was extracted from the ticks and from cultured spirochaetes. Sequence analysis of the flagellin gene revealed that the DNA obtained was from B. garinii, regardless of the geographical origin of the sample. Identical fla gene fragments in ticks collected in both hemispheres indicate a transhemispheric exchange of B. garinii. A marine ecological niche and epidemiological route for Lyme disease Borrelia are proposed. The prevalence of B. burgdorferi s.l. infected ticks on migrating passerine birds was studied. A total of 22, 998 birds were caught and examined for ticks. The presence of spirochaetes in the 967 collected ticks was determined by DNA amplification by PCR on all ticks. To determine which B. burgdorferi s.l. species were present, classification was performed by DNA amplification using species-specific 16S rDNA primers and by DNA sequencing. Flagellin gene sequences of all species of B. burgdorferi s.l. previously recorded in Europe were found. B. garinii was the most prevalent. These data support the notion that passerine birds are at least partly responsible for the distribution of Lyme disease Borrelia spirochaetes in Europe. To elucidate the distribution of B. burgdorferi s.l. in subarctic regions, strains isolated from I. ricinus and I. uriae ticks found on islands in the northern Atlantic and Baltic Sea were characterised molecularly. All isolates were verified as B. garinii by 16S-rRNA gene analysis and immunoblotting using monoclonal antibodies specific for the outer surface proteins A and C. Three ribotypes (RT's) of B. garinii were found. The I. ricinus associated RT1 is phenotypically the most heterogeneous. RT2 is restricted to the islands in the northern Baltic Sea, whereas RT3 was also recovered from ticks found on islands in the North Atlantic. The heterogeneity of the B. garinii population in the Baltic Sea might be influenced by two geographically opposite directions, North Atlantic (RT3) and Euroasia (RT1). / digitalisering@umu.se

The aim of present master’s thesis was to compare the results of serological methods for diagnosing borreliosis that are commonly used in Spadia laboratories (ELISA, immunoblots) in terms of recommendation on how and when to indicate and interpret them. The theoretical part is focusing on the characteristics and history of borreliosis, microbiological description of Borrelia, immune system and pathogenesis of the disease as well as the therapy and prevention. The experimental part is focusing on the analysis of results obtained from common examinations of antibodies to Borrelia made in Spadia Lab laboratories from January 1st 2014 to December 31st 2015. Screening of antibodies to Borrelia made by ELISA in IgM and IgG was done for all samples according to recommendation of CDC. In 2014 the ELISA screening was done using ELISA kits from Euroimmun and Evolis sample processors whereas in 2015 it was done using DiaSorin’s CLIA kits on Liaison analyser. Positive results were then confirmed by Westernblot or lineblot alternatively if the physician did not ask otherwise. It must be remembered that ELISA and Westernblot belong among serological methods that are using antibodies, i.e. substances produced by the immune system. The immune system plays the key role in protecting the body against infection and the antibodies are its important tool. Serological methods belong among immunoassay methods, which is still not standardized. Diagnosis of infections cann‘t be based only on antibody testing. It is necessary to assess the results in the context of the entire clinical picture, history and in the case of antibodies it is recommended retesting with an interval.