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In recent years, the distribution of Dermacentor reticulatus ticks has expanded into new territories in many European countries, including Poland, with increased population densities in areas of their regular occurrence. The spread of D. reticulatus enhances the risk of exposure of domestic animals and their owners to tick-borne diseases. The objective of this study was to assess the prevalence of infection of D. reticulatus ticks feeding on dogs with the pathogens Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum. The study material comprised 152 D. reticulatus ticks collected from dogs in the northeastern part of Lublin Province (eastern Poland). A ready-made AmpliSens® TBEV, B.burgdorferi sl, A.phagocytophilum, E.chaffeensis/E.muris-FRT PCR kit was used for qualitative detection and differentiation of tick-borne infections. The assessment of the degree of infection of the analyzed ticks with the two pathogens revealed that 9.2% (14/152) of the examined ticks were infected with one of the pathogens. No co-infections with the pathogens were detected in any of the ticks. The highest specific percentage of infections (8.6%, 13/152) was associated with A. phagocytophilum. The presence of B. burgdorferi s.l. was detected in only one of the examined ticks (0.7%). The spread of D. reticulatus to new territories and the increase in population density in areas of their regular occurrence implies the need for further studies of the prevalence of pathogens with medical and veterinary importance in order to assess the risk of tick-borne diseases.

The aim of the study was to analyze the concentrations of some serological parameters in patients undergoing antibiotic therapy for LD, and to assess the potential use of IgG anti-C6VlsE in the assessment of treatment effectiveness.The study involved 21 patients diagnosed with LD. To determine the levels of anti-C6VlsE IgG, metalloproteinases and cytokines, blood was collected three times: before treatment, immediately after treatment, and 3-4 months after treatment.Immediately after the antibiotic therapy, all the patients in the study group declared improvement in their health. At 3-4 months post-treatment, 50% of patients reported recurrence of the disease symptoms. The anti-C6VlsE IgG concentrations at the analyzed time points exhibited varied dynamics: in 15% of patients, a decrease in the concentration of anti-C6VlsE was observed in the three consecutive serum samples; an increase of anti-C6VlsE IgG was detected in 35% of cases; in 15% of the subjects, the concentration of anti-C6VlsE IgG increased immediately after the antibiotic therapy and declined after 3-4 months, whereas a reverse situation was noted in 10% of the patients.The presence of high concentrations of anti-C6VlsE IgG is associated with appearance of response to Borreliella spp. infection, which suggests that the parameter can be used in the diagnosis of the active form of borreliosis. However, from a practical point of view, a marker of successful elimination of infection should indicate the effectiveness or failure of antibiotic therapy within a relatively short post-treatment period. The anti-C6VlsE IgG antibodies do not seem to satisfy this provision.

OBJECTIVES The aim of the study was to analyze some metalloproteinases, cytokines, and chemokines in LB patients and healthy seropositive subjects. The presence of IgM/IgG antibodies against specific Borreliella antigens was analyzed in the presence or absence of clinical manifestations of LB. MATERIAL AND METHODS The study involved 38 patients diagnosed with LB and arthralgia and/or arthritis symptoms, and 57 foresters presenting no clinical symptoms of LB. The ELISA test was applied for general screening of anti-Borreliella IgM/IgG. Western blot was used for confirmatory diagnosis of LB for the positive and borderline results. Serum IL-2, IL-4, IL-6, IL-10, IL-17A, IFN-γ, TNF, IL-8, CCL5, CXCL9/MIG, CCL2/MCP-1, CXCL10/IP-10 concentrations were measured with the use of the Human Cytometric CBA test. The concentration of MMP-2 and MMP-9 in the serum was determined with the use of ELISA tests. RESULTS Analysis of the cytokines and chemokines revealed that only the concentration of IL-2 was significantly higher (2.4 pg/m; p=0.00641) in patients with LB symptoms than in the seropositive individuals (0.4 pg/ml). The MMP2 concentration was significantly higher (233.3 ng/ml; p=0.00294) in patients with clinical manifestations of LB than in those occupationally exposed to tick bites, but did not have anti-Borreliella antibodies (192.0 ng/ml). CONCLUSIONS The presence of IgG antibodies against a number of Borreliella antigens and the differences in the IL-2 and MMP2 levels in seropositive or seronegative individuals and symptomatic LB patients, may indicate differences in the intensity of the immune response to the infection and, consequently, may induce development of clinical manifestations of the disease in seropositive and seronegative individuals.

BACKGROUND This study is aimed at evaluating the awareness among hunters and foresters concerning the activities undertaken for the prevention of Borrelia burgdorferi infections. MATERIAL AND METHODS The study group included 236 hunters from Poland and 339 foresters from Ukraine. The study was based on an original questionnaire including questions which concerned the applied methods of removing ticks attached to the skin, the kind of preventive measures undertaken to avoid tick bites, the evaluation of knowledge concerning the prevention of tick-borne diseases, as well as the participants' willingness to be educated on this issue. RESULTS Removing a tick with a pair of tweezers by a simple vertical move was more frequently practiced by the hunters (16.5%). Spreading an oily substance over a tick to make it go out on its own was more often applied by the foresters (18.8%). The occurrence of erythema migrans was more frequently reported by the foresters (47.2%). Among the persons declaring no knowledge of the prevention of tick-borne diseases, 77.8% of the hunters and 61.5% of the foresters were willing to gain such knowledge, and 11.1% of the hunters and 21.2% of the foresters did not want any education on this issue. The hunters more often than the foresters applied repellents and checked their body after returning from green areas. CONCLUSIONS Education is needed in certain groups undertaking hobby or vocational activities in forest areas of Poland and Ukraine. It seems reasonable to undertake research on the preferences concerning the choice of repellents among the groups at risk of tick bites. Med Pr. 2020;71(1):59-68.

Background: Infections in pregnant women or women planning pregnancy caused by the protozoan Toxoplasma gondii and the viruses varicella zoster virus (VZV) and human parvovirus B19 can be a danger to the fetus. Aim of the study: The aim of the study was to determine the serological status of women of childbearing age in relation to T. gondii, VZV and human parvovirus B19 in a region of Eastern Poland (Biała Podlaska District). Material and methods: The study group consisted of 174 women aged 19 to 35 (average 23, SD 3.68) from the Biała Podlaska District. Anti-T. gondii IgM/IgG antibodies, anti-VZV IgG and anti-human parvovirus B19 IgG were detected by ELISA. Results: Serological screening revealed that the most common antibodies were anti-VZV and anti-parvovirus B19 (in 96% and 60.9% of women, respectively). Anti-T. gondii antibodies were found in 28.6%. No correlation was found between the presence of anti-T. gondii, human parvovirus B19, and VZV antibodies and the age of the examined women, their place of residence, and their education. Conclusions: About 4%, 39% and 71.2% of women participating in this study were still susceptible to infection with VZV, human parvovirus B19, and T. gondii, respectively. It is therefore important to address health education primarily in women of childbearing age in order to help them undertake relevant measures for prevention of T. gondii, human parvovirus B19 and VZV infection.

Ixodes ricinus followed by Dermacentor reticulatus are the most common tick species with medical and veterinary importance in Poland. Ixodes ricinus is widely distributed across Poland, whereas the range of D. reticulatus is limited to the eastern and central parts of the country, with several new foci in the mid-west and west. Concurrently, an increase in the occurrence range of D. reticulatus has been observed. Recent studies indicate its high density in eastern Poland. The aim of the study was to determine the species composition of ticks infesting dogs from the northeastern part of Lublin Province. In a three-year study (2017-2019), 501 tick specimens were collected from 122 dogs in the north-eastern part of Lublin Province (eastern Poland). 174 ticks were removed from 24 pet dogs kept by individual owners and the other 327 were collected from 98 dogs from the animal shelter. Two tick species: D. reticulatus 55.5% (278/501) and I. ricinus 44.5% (223/501) were identified in the group of 501 ticks removed from the dogs. Dogs in the north-eastern part of Lublin Province are exposed to contact with two tick species: I. ricinus and D. reticulatus, with a higher frequency of the latter species. Identification of the species of ticks infesting dogs in an analyzed area and knowledge of pathogens transmitted by ticks facilitates correct differential diagnosis of tick-borne diseases.

The aim of the study was to analyze the selected nanomechanical properties of Candida spp: Candida albicans (standard strain ATCC 10231), Candida albicans (clinical strain, cultured from an oral swab), Candida lipolytica (clinical strain, cultured from a nosal swab) in atomic force microscopy (AFM). The culture Candida spp. was performed of Tryptone Soya Broth (BioMaxima). The topography and sample properties were analysed in AFM (Ntegra Spectra C from NT) and the results were carried out using NOVA 1.1.0.1824 software. C. albicans ATCC 10231 cells were significantly higher 1.81 μm (p = 0.001) from clinical strains: C. albicans (1.30 μm) and C. lipolytica (1.23 μm). C. albicans ATCC 10231 cells, and C. albicans cells of the clinical strain were softer, especially in the top parts of cells, than C. lipolytica cells. Adhesion force measured for C. albicans ATCC 10231 was 62.83 nN, and was significantly higher compared to the values obtained for C. albicans (41.93 nN, p = 0.0002 ) and C. lipolytica (41.78 nN, p = 0.0002 ). The stiffness of the Candida spp. cell surface was comparable and was in the range of 5–6 nA. The differences in height may result from different conditions in which clinical strains grow. Adhesion force can be helpful in the analysis of the degree of destruction of the cell wall by various substances. The conducted analyses showed morphological differences and the differences in mechanical properties of the researched Candida spp. This data may be important in assessing their susceptibility to the effects of various substances of a lytic nature.

Tick-borne diseases (TBD) are an increasing health problem for humans, pets, and livestock. It is closely related to the increasing frequency of ticks. The aim of the study was to identify Borrelia burgdorferi, Anaplasma phagocytophilum, Francisella tularensis, and Coxiella burnetii in ticks from selected regions of north-eastern Poland. The material for molecular tests were ticks belonging to two species: Dermacentor reticulatus (meadow tick) and Ixodes ricinus (common tick), collected in the period from July 2020 to June 2021. The studies were performed on arachnids collected mostly from the ground (90.4%), as well as removed from dogs (8.4%) and humans (1.2%). The genetic material was amplified by PCR in the multiplex PCR variant (B. burgdorferi and A. phagocytophilum) and single PCR (F. tularensis, C. burnetii). Only 9 out of 500 ticks tested showed genetic material indicating infection with the above-mentioned microorganisms (1.8%). D. reticulatus ticks dominated in this group. The highest number of ticks, that is seven D. reticulatus and one I. ricinus, were infected with B. burgdorferi. Only one I. ricinus tick (0.3%) and one D. reticulatus tick (0.2%) carried genetic material of A. phagocytophilum. In the latter case, it was a co-infection with B. burgdorferi. Tests on ticks showed a low degree of infection with B. burgdorferi and A. phagocytophilum bacteria (1.8%) and no genetic material of F. tularensis or C. burnetii. On the other hand, one D. reticulatus tick was coinfected with B. burgdorferi and A. phagocytophilum (0.2%). The low percentage of infected I. ricinus ticks may have been due to the small number of specimens examined, which in turn resulted from the fact that D. reticulatus was the dominant tick species in the research area.

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 aim of our study was the collection of seroprevalence data for Toxoplasma gondii, Coxiella burnetii, Trichinella spp., and Francisella tularensis from hunters in Lublin Province. The antibodies against T. gondii and C. burnetii were recorded in 38.5% and 16.2% of the sera, respectively. 4.05% of the sera were seropositive for both T. gondii and C. burnetii. None of the sera tested reacted positively with F. tulariensis or Trichinella spp. Seroprevalence of T. gondii and C. burnetii is common among the hunters from Lublin Province. It seems reasonable to undertake similar research among hunters from other regions of eastern Poland.

Introduction: Although HEV infection in pigs does not pose a major economic risk to pork production, the risk of zoonotic transmission to humans is an important aspect of public health. HEV genotype 3 infections were reported in developed countries in individuals who had consumed raw meat or meat products from deer, wild boars, or pigs. The aim of the study was the analysis of the occurrence of HEV-specific antibodies among wild boars and domestic pigs in Poland. Material and Methods: A total of 290 samples from wild boars and 143 samples from pigs were tested. The antibodies were tested by ELISA. Results: The presence of anti-HEV IgG was demonstrated in 44.1% of pigs and 31.0% of wild boars. Anti-HEV IgG antibodies were detected in 1.4% of samples from pigs and in 2.1% of samples from wild boars at borderline level. The statistical analysis shows significant differences in the positive results for anti-HEV IgG between the groups of pigs and wild boars (P = 0.0263). Conclusion: Regular surveillance of the occurrence of HEV in swine and wild boars should be performed in the future.

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.

International audience; Small molecules containing quinone and/or oxime moieties have been found as promising anti-fungal agents. One of them is 4-AN, a recently reported potent anti-Candida compound, which inhibits the formation of hyphae, decreases the level of cellular phosphoproteome, and finally shows no toxicity towards human erythrocytes and zebrafish embryos. Here, further research on 4-AN is presented. The results revealed that the compound: (i) Kills Candida clinical isolates, including these with developed antibiotic resistance, (ii) affects mature biofilm, and (iii) moderately disrupts membrane permeability. Atomic force microscopy studies revealed a slight influence of 4-AN on the cell surface architecture. 4-AN was also shown to inhibit multiple various protein kinases, a characteristic shared by most of the ATP-competitive inhibitors. The presented compound can be used in novel strategies in the fight against candidiasis, and reversible protein phosphorylation should be taken into consideration as a target in designing these strategies.

Background Atomic force microscopy (AFM) is an experimental technique which recently has been used in biology, microbiology, and medicine to investigate the topography of surfaces and in the evaluation of mechanical properties of cells. The aim of this study was to evaluate the influence of the complement system and specific anti-Borrelia antibodies in in vitro conditions on the modification of nanomechanical features of B. burgdorferi B31 cells. Material and methods In order to assess the influence of the complement system and anti-Borrelia antibodies on B. burgdorferi s.s. B31 spirochetes, the bacteria were incubated together with plasma of identified status. The samples were applied on the surface of mica disks. Young's modulus and adhesive forces were analyzed with a NanoScope V, MultiMode 8 AFM microscope (Bruker) by the PeakForce QNM technique in air using NanoScope Analysis 1.40 software (Bruker). Results/conclusion The average value of flexibility of spirochetes' surface expressed by Young's modulus was 10185.32 MPa, whereas the adhesion force was 3.68 nN. AFM is a modern tool with a broad spectrum of observational and measurement abilities. Young's modulus and the adhesion force can be treated as parameters in the evaluation of intensity and changes which take place in pathogenic microorganisms under the influence of various lytic factors. The visualization of the changes in association with nanomechanical features provides a realistic portrayal of the lytic abilities of the elements of the innate and adaptive human immune system.

Sustainable technologies are increasingly promoted in various production areas. Protection of natural resources, as well as rational waste management, may lead to better optimization of technologies. Biochar, a product of pyrolysis of organic residues has found wide applications in waste management, agriculture, energy and construction industry. In the present study biochar samples produced in Poland and in Brazil were analysed for microbial content using three substrates: Plate Count Agar, Malt Agar, and Potato Agar. Both qualitative and quantitative measurements were done. Microscopic analysis of the biochar structure was also performed. We found that microbial cultures in both biochars represented a wide range of biodiversity of microorganisms genera and species. We demonstrate that the biochar samples differ depending on the botanical origin as well as on the production technology. Structure of the tested samples also varied depending on the botanical origin. Sample 1-PL (pine) was characterised by a compact and regular structure, while sample 2-PL (oak) showed porous and irregular structure. Sample from Brazil (1-BR) showed a more delicate structure than Polish biochars. Obtained properties may suggest a range of implications for practice.

Tick-borne diseases are infectious diseases transmitted by the bites of infected ticks. The most common vector-borne disease in Europe is Lyme borreliosis (LB). The number of the reported cases of LB has been steadily increasing. For this reason, it is important to undertake educational activities in the field of tick-borne diseases. The aim of the study was to investigate the level and scope of knowledge on tick-borne diseases among nursing students from Poland and Slovakia. The study sample consisted of 428 nursing students (192 students of the State School of Higher Education in Biała Podlaska, Poland and 236 students of the University in Presov, Slovakia). The knowledge of the surveyed students on tick-borne diseases is limited. Polish students demonstrated greater knowledge about LB, while students from Slovakia showed greater awareness of the threat posed by tick-borne encephalitis. Particularly worrying is the lack of knowledge on proper way of removal of a tick attached to the skin. Students recognize the need to broaden their knowledge on tick-borne diseases, which should be taken into consideration in realized studies curricula.

Purpose. In this study, we applied various assays to find new activities of 1,4-naphthoquinone derivatives for potential anti-Candida albicans applications. Methodology. These assays determined (a) the antimicrobial effect on growth/cell multiplication in fungal cultures, (b) the effect on formation of hyphae and biofilm, (c) the influence on cell membrane integrity, (d) the effect on cell morphology using atomic force microscopy, and (e) toxicity against zebrafish embryos. We have demonstrated the activity of these compounds against different Candida species and clinical isolates of C. albicans. Key findings. 1,4-Naphthoquinones significantly affected fungal strains at 8–250 mg l−1 of MIC. Interestingly, at concentrations below MICs, the chemicals showed effectiveness in inhibition of hyphal formation and cell aggregation in Candida. Of note, atomic force microscopy (AFM) analysis revealed an influence of the compounds on cell morphological properties. However, at low concentrations (0.8–31.2 mg l−1), it did not exert any evident toxic effects on zebrafish embryos. Conclusions. Our research has evidenced the effectiveness of 1,4-naphthoquinones as potential anti-Candida agents.