Tick exposure and prevalence of Borrelia burgdorferi antibodies among hunters and other individuals exposed to vector ticks in eastern Poland

Background: Lyme borreliosis is the most frequent tick-borne disease in Europe and North America, and the number of registered cases is on the increase. Frequent presence in the habitats of ticks enhances the risk of tick bites and possible infection with Borrelia burgdorferi spirochetes. Objective: The aim of the study was to assess the risk of B. burgdorferi infection posed to hunters and other individuals exposed to activity-related contact with ticks. Material and methods: The study was carried out in the northern part of the Lublin Province (eastern Poland) and involved 150 individuals exposed to tick bites (110 hunters and 40 individuals exposed to activity-related contact with ticks). The analysis of sera for the presence of B. burgdorferi IgM and IgG antibodies was carried out. All 150 individuals were tested with the ELISA assay, and positive and borderline results of the assay were verified with the Western blot test. All study participants completed a questionnaire, which provided information about exposure to ticks, application of prophylactic measures, and awareness of Lyme borreliosis. Results: The ELISA assay revealed a positive or borderline result in at least one of the classes of B. burgdorferi antibodies in 63.3% (95/150) of the individuals (IgM 14.0%, IgG 63.3%). Verification carried out with the Western blot test showed a positive or borderline result in at least one of the antibody classes in 38.0% (57/150) of the examined persons (IgM 2.7%, IgG 36.7%). Abdomen (56.0%) and legs (53.7%) were the most frequently bitten body regions. Tick bites on the abdomen were significantly more frequently declared by hunters. Inspection of the body after returning from natural areas was more popular prophylactic method than use of repellents. Inspection of the body was significantly more often used in the group of the hunters. Conclusions: The risk of B. burgdorferi infection among hunters and other individuals undertaking activities associated with exposure to tick bites in the study area is high.

Prevalence of Borrelia Burgdorferi Sensu Lato in Ticks from the Ternopil Region in Ukraine.

INTRODUCTION: Lyme borreliosis/Lyme disease is caused by Borrelia burgdorferi and is one of the most common vector-borne diseases transmitted by ticks. MATERIAL AND METHODS: A total of 136 Ixodes ricinus ticks, collected in the Ternopil (Ukraine) region, including 126 adults (70 females and 56 males), and 10 nymphs were examined. The identification of the species and their developmental form was based on morphological characteristics. RESULTS: PCR with B5S-Bor and 23S-Bor primers resulted in Borrelia burgdorferi sensu lato DNA amplification among six ticks (4.4%). The detailed analysis based on the DNA sequencing showed the presence of DNA of Borrelia afzelii in four samples; the remaining two represented Borrelia burgdorferi sensu lato complex, although their genospecies were not determined. The research confirmed the dominance of Borrelia afzelii genospecies in the ticks from Ukraine. CONCLUSION: It seems reasonable to undertake similar research in ticks from other regions of Ukraine. Knowledge in this field can be useful for public health and planning the prevention of tick-borne diseases.

The Serological Surveillance of Hepatitis E virus among Hunters and Foresters in Eastern Poland.

The aim of the study was the analysis of the occurrence of specific antibodies against HEV among hunters and foresters who are at risk to be exposed. The study group consisted of 210 hunters (23-80 years old) and 60 foresters (22-64 years old). Anti-HEV IgG were present in 3.81% of the samples of the hunters and in 5% of the samples of the foresters. The statistical analysis shows no significant differences in the results anti-HEV IgG between the groups of hunters and foresters (p = 0.5278). Significantly higher anti-HEV IgG titers were found in the older age group (> 55 years old).

Seroprevalence of Rubella and Cytomegalia in Young Women from Biala Podlaska District.

The aim of this study was to analyze the seroprevalence of rubella and cytomegalia among young women. The study included 175 healthy women from the Biala Podlaska District, aged 16 to 35 years. Anti-rubella and anti-CMV IgG were determined by ELISA. 172 (98.3%) study subjects tested positive for rubella antibodies, 1 (0.6%) was seroindeterminate and 2 (1.1%) were seronegative. CMV antibodies were detected in 119 (68.0%) participants; the series included also 1 (0.6%) seroindeterminate and 55 (31.4%) seronegative women. The levels of rubella and CMV antibodies were not significantly affected by age, place of residence and educational level of the study subjects.

The incidence of Borrelia burgdorferi, Anaplasma phagocytophilum and Babesia microti coinfections among foresters and farmers in eastern Poland.

BACKGROUND & OBJECTIVES: Lyme borreliosis is the most common tick-borne disease in Europe and the USA. However, a great variety of pathogens are transmitted by ticks, which results in mixed infections, with Lyme borreliosis. The aim of the present study was to show the incidence of Borrelia burgdorferi, Anaplasma phagocytophilum, and Babesia microti coinfections among the population of foresters and farmers, as these people, due to their profession, are particularly exposed to tick contact. METHODS: The study was carried out in eastern Poland (the northern part of the Lublin Province) in 2013. The study was performed in a group of 93 individuals occupationally exposed to tick bites (foresters and farmers), whose blood serum showed the presence of IgG anti-B. burgdorferi antibodies. Blood serum in this group were evaluated for IgG anti-A. phagocytophilum and IgG anti-B. microti antibodies by means of IFA IgG indirect immunofluorescence tests. Information related to age, sex, number of tick bite episodes, presence of various symptoms related to the tick bites, and antibiotic therapy applied as treatment for diagnosed Lyme borreliosis were obtained from the subjects through a structured questionnaire. The results were analyzed in Statistica v. 7.1 statistical analysis software. RESULTS: The presence of IgG antibodies against the analyzed pathogens revealed B. burgdorferi and A. phagocytophilum coinfection in 26 (28%) of the examined subjects and B. burgdorferi and B. microti coinfection in one person (1.1%). No coinfection with all the three pathogens was observed in any individual. The co-occurrence of headache plus bone, joint and muscle pain was noted significantly more often among individuals diagnosed with B. burgdorferi and A. phagocytophilum coinfection. INTERPRETATION & CONCLUSION: Foresters and farmers are exposed to B. burgdorferi and A. phagocytophilum coinfection in the study area. Therefore, it is probable that these pathogens may severely interfere with the clinical course of Lyme borreliosis.

Serological surveillance of vector-borne and zoonotic diseases among hunters in eastern Poland.

BACKGROUND & OBJECTIVES: Many etiological agents of zoonoses are considered as significant biological hazard to people visiting forested areas frequently, for instance, hunters. They may be exposed to ticks, rodents, and birds as well as excreta/secretions of wild animals or contaminated water and soil. Hence, this population is at risk of contracting infection with pathogens such as Borrelia burgdorferi sensu lato (s.l.), Anaplasma phagocytophilum, Babesia spp., tick-borne encephalitis virus, Bartonella spp., Francisella tularensis, Echinococcus spp., or hantaviruses. The aim of the study was to assess the seroprevalence of zoonotic agents, viz. A. phagocytophilum, hantaviruses, and Echinococcus spp., with special regard to B. burgdorferi s.l., among hunters in Lubelskie Voivodeship (eastern Poland). METHODS: Serum samples collected from 134 hunters from Lubelskie Voivodeship were analyzed with the use of immunological techniques (enzyme-linked immunosorbent assay, line immunoblot test, and indirect fluorescence assay) for the presence of antibodies against the agents. RESULTS: Specific antibodies were detected in 66% of the tested individuals. Antibodies against B. burgdorferi s.l. (39%), A. phagocytophilum (30%), hantaviruses (9%), and Echinococcus spp. (8%) were detected individually or as mixed results. INTERPRETATION & CONCLUSION: The results confirm that there is a risk of exposure to different pathogens in the forested areas in eastern Poland and that hunters are highly vulnerable to infection with the examined zoonotic agents. A significant proportion of co-occurring antibodies against different pathogens was noticed. Thus, hunters have to take special care of their health status evaluation and mitigate the exposure risk by using adequate prophylaxis measures.

[Monoinfections caused by Borrelia burgdorferi and Borrelia burgdorferi / Anaplasma phagocytophilum co-infections in forestry workers and farmers]. / Monoinfekcja Borrelia burgdorferi i wspólzakazenia Borrelia burgdorferi / Anaplasma phagocytophilum u pracowników lesnictwa i rolników.

BACKGROUND: The presence of co-infections induced by tick-borne pathogens in humans is an important epidemiological phenomenon. This issue has attracted growing attention of doctors and people working under conditions of an increased risk of being exposed to tick bites. MATERIAL AND METHODS: The research group consisted of 93 individuals with current anti-immunoglobulin M/G (IgM/ IgG) Borrelia burgdorferi or IgG anti-Anaplasma phagocytophilum. The respondents were identified during the screening survey in a group of farmers and foresters occupationally exposed to tick bites. The aim of the work was to analyse the frequency of antibodies to specific antigens of B. burgdorferi and the levels of cytokines in forestry workers and farmers with B. burgdorferi monoinfections and B. burgdorferi / A. phagocytophilum co-infections. Statistical analysis was performed using the Chi2, Mann-Whitney U and Kruskal-Wallis tests. RESULTS: There is a stronger generation of IgG antibodies to B. burgdorferi antigens in patients with B. burgdorferi / A. phagocytophilum co-infections, such as variable major protein-like sequence expressed (VlsE) (p < 0.05), p19 (p < 0.02), p17 (p < 0.05) and complement regulator-acquiring surface protein 3 (CRASP3) (p < 0.02) compared to persons with B. burgdorferi monoinfections. The discrepancies in the synthesis of cytokines interleukin 6 (IL-6), IL-10, and tumor necrosis factor α (TNF-α) have not been found in persons with B. burgdorferi monoinfections and B. burgdorferi / A. phagocytophilum co-infection. CONCLUSIONS: The immune response directed against B. burgdorferi is stronger in patients co-infected with B. burgdorferi and A. phagocytophilum than in those with monoinfection.

The role of programmed death ligand 1 pathway in persistent biomaterial-associated infections.

Staphylococcus epidermidis is commonly involved in biomaterial-associated infections. Bacterial small colony variants (SCV) seem to be well adapted to persist intracellularly in professional phagocytes evading the host immune response. We studied the expression of PD-L1/L2 on macrophages infected with clinical isolates of S. epidermidis SCV and their parent wild type (WT) strains. The cytokine pattern which is triggered by the examined strains was also analysed. In the study, we infected macrophages with S. epidermidis WT and SCV strains. Persistence and release from macrophages were monitored via lysostaphin protection assays. Moreover, the effect of IFN-γ pre-treatment on bacterial internalisation was investigated. Expression of PD-L1/L2 molecules was analysed with the use of FACS. Inflammatory reaction was measured by IL-10, TNF-α ELISAs, and transcriptional induction of TNF-α. Our study revealed that clinical SCV isolates were able to persist and survive in macrophages for at least 3 days with a low cytotoxic effect and a reduced proinflammatory response as compared to WT strains. Bacteria upregulated PD-L1/L2 expression on macrophages as compared to non-stimulated cells. The results demonstrated that the ability of S. epidermidis SCVs to induce elevated levels of anti-inflammatory cytokine, IL-10, and reduced transcriptional induction of TNF-α, together with expression of PD-L1 on macrophages and the ability to persist intracellularly without damaging the host cell could be the key factor contributing to chronicity of SCV infections.

[Risk of occupational infections caused by Borrelia burgdorferi among forestry workers and farmers]. / Ryzyko zakazen zawodowych Borrelia burgdorferi u pracowników lesnictwa i rolników.

BACKGROUND: The aim of the work was to analyze the incidence of infection with Borrelia burgdorferi in forestry workers and farmers, major groups occupationally exposed to tick bites. MATERIAL AND METHODS: The study group included 275 workers (171 foresters and 104 farmers). The control group consisted of 45 people, who have not been occupationally exposed to tick bites. The screening Elisa and Wb tests for the presence of anti-Borrelia IgM/IgG antibodies were performed in all subjects of the study and control groups. Statistical analysis was performed using the Chi2 test. RESULTS: The positive results denoting the presence of anti-Borrelia IgM/IgG antibodies were found in 55% of farmers and 28% of foresters occupationally exposed to Lyme borreliosis and coming from the area of South Podlasie Lowland and Lublin Polesie. The differences between the forestry workers and the control group (p < or = 0.00001) and between farmers and the control group (p < or = 0.001) were statistically significant. The species, such as B. spielmanii and B. bavariensis, which have not yet been reported in Poland, are significant etiologic agents of Lyme disease. CONCLUSION: The risk of occupational exposure to the B. burgdorferi infection is high for foresters and farmers, and the infection with spirochetes is frequently confirmed on the basis of positive results of the Wb test. The presence of specific antibodies against protein antigens of B. spielmanii and B. bavariensis suggest that these bacteria can cause Lyme disease both independently and in participation with other Borrelia species, which influences the development of the clinical manifestations of infection.

Characterization of Staphylococcus epidermidis and Staphyloccocus warneri small-colony variants associated with prosthetic-joint infections.

We determined the frequency of isolation of staphylococcal small-colony variants (SCVs) from 31 culture-positive patients undergoing revision of total hip prosthesis for aseptic loosening or presumed prosthetic-joint infection (PJI). We analysed auxotrophy of cultured SCVs, their antimicrobial susceptibility profiles and their biofilm-forming capacity. Eight SCV strains were cultivated from six (19 %) patients. All SCVs were coagulase-negative staphylococci (CNS) with Staphylococcus epidermidis as the predominant species; there was also one Staphylococcus warneri SCV. The SCVs were auxotrophic for haemin, with one strain additionally auxotrophic for menadione. We noted the presence of two phenotypically (differences concerning antimicrobial susceptibility) and genetically distinct SCV strains in one patient, as well as the growth of two genetically related SCVs that differed in terms of their morphology and the type of auxotrophy in another. Seven out of eight SCVs were resistant to meticillin and gentamicin. In addition, antibiotic sensitivity testing revealed three multidrug-resistant SCV-normal-morphology isolate pairs. One S. epidermidis SCV harboured icaADBC genes and was found to be a proficient biofilm producer. This paper highlights the involvement of CNS SCVs in the aetiology of PJIs, including what is believed to be the first report of a S. warneri SCV. These subpopulations must be actively sought in the routine diagnosis of implant-associated infections. Moreover, in view of the phenotypic and genetic diversity of some SCV pairs, particular attention should be paid to the investigation of all types of observed colony morphologies, and isolates should be subjected to antimicrobial susceptibility testing.

[Significance of circulating immune complexes (CIC) in the diagnosis of infections with Borrelia burgdorferi]. / Znaczenie krazacych kompleksów immunologicznych (CIC) w diagnostyce zakazen Borrelia burgdorferi.

INTRODUCTION: The infection caused by Borrelia burgdorferi initiates a number of reactions in the human immunological system and particular behaviors of the infectious factor. The aim of work was to analyze the level of circulating immune complexes (CIC) due to infection with Borrelia burgdorferi in persons professionally exposed to tick bites. MATERIAL AND METHODS: The group of tested consisted of 275 persons, who are professionally exposed to tick bites, including 171 foresters and 104agriculturists. The controlled group consisted of 45 people, living permanently ortemporarily in Biata Podlaska, city in the South Podlasie Lowland, who have not been exposed professionally on tick bites.The screening test ELISA (Euroimmun) has been carried among all persons from the tested group towards the presence ofantibodies IgM/IgG anty-Borrelia. Among those who met positive or delimited results from screening test Elisa, another test of Western blot (Wb, Euroimmun, Virotech GmbH) has been carried in order to confirm the infection. The serum which has been taken from a group of persons being professionally exposed to infection of B. burgdorferi and persons from a controlled group, has been tested in terms of estimating levels of circulating immune complexes CIC C1q (Elisa, DRG). RESULTS AND CONCLUSION: The circulating immune complexes (CIC) are increasingly generated at persons with present IgM and/or IgG anty-Borrelia but their existence accompanied by the presence of strongly expressed humoral response does not affect the diagnostic effectiveness of infections with B. burgdorferi.