Prevalence of Borrelia, Neoehrlichia mikurensis and Babesia in ticks collected from vegetation in eastern Poland.

In Poland, tick-borne diseases constitute the majority of diseases related to exposure to biological agents with a predominance of Lyme borreliosis; therefore, research on ticks as a reservoir of various pathogens remains crucial in the epidemiology of human diseases after tick bites. This study aimed to identify the occurrence of Borrelia burgdorferi sensu lato, Borrelia miyamotoi, Neoehrlichia mikurensis, and Babesia spp. in ticks collected from vegetation in eastern Poland. Additionally, the prevalence of co-infections in the adult Ixodes ricinus ticks was determined. Among I. ricinus ticks the predominantly detected pathogen was B. burgdorferi s.l. (23%) with B. burgdorferi sensu stricto as the most frequently identified species, followed by B. garinii. In 2013, the double or triple infections of B. burgdorferi s.s., B. afzelii, and B. garinii species did not exceed 9% in adult ticks, whereas in 2016, the prevalence of mixed infections reached 29%. The prevalence of N. mikurensis and B. miyamotoi in I. ricinus was determined at the same level of 2.8%. Four Babesia species were identified in the examined I. ricinus population: B. microti (1.5%), B. venatorum (1.2%), B. divergens (0.2%), and B. capreoli (0.1%). Co-infections were detected in 10.1% of all infected ticks with the highest prevalence of co-infections with B. burgdorferi s.l. and Babesia species. The changes in the prevalence and the distribution of particular pathogens within tick populations indicate the need for monitoring the current situation related to tick-borne pathogens from the aspect of risk to human health.

Cathelicidin Treatment Silences Epithelial-Mesenchymal Transition Involved in Pulmonary Fibrosis in a Murine Model of Hypersensitivity Pneumonitis.

Pulmonary fibrosis is becoming an increasingly common pathology worldwide. Unfortunately, this disorder is characterized by a bad prognosis: no treatment is known, and the survival rate is dramatically low. One of the most frequent reasons for pulmonary fibrosis is hypersensitivity pneumonitis (HP). As the main mechanism of pulmonary fibrosis is a pathology of the repair of wounded pulmonary epithelium with a pivotal role in epithelial-mesenchymal transition (EMT), we assumed that EMT silencing could prevent disease development. Because of several biological features including wound healing promotion, an ideal candidate for use in the treatment of pulmonary fibrosis seems to be cathelicidin. The aim of the studies was to understand the influence of cathelicidin on the EMT process occurring during lung fibrosis development in the course of HP. Cathelicidin's impact on EMT was examined in a murine model of HP, wherein lung fibrosis was induced by chronic exposure to extract of Pantoea agglomerans (SE-PA) by real-time PCR and Western blotting. Studies revealed that mouse exposure to cathelicidin did not cause any side changes in the expression of investigated genes/proteins. Simultaneously, cathelicidin administered together or after SE-PA decreased the elevated level of myofibroblast markers (Acta2/α-smooth muscle actin, Cdh2/N-cadherin, Fn1/Fibronectin, Vim/vimentin) and increased the lowered level of epithelial markers (Cdh1/E-cadherin, Ocln/occludin). Cathelicidin provided with SE-PA or after cessation of SE-PA inhalations reduced the expression of EMT-associated factors (Ctnnd1/ß-catenin, Nfkb1/NFκB, Snail1/Snail, Tgfb1/TGFß1 Zeb1/ZEB1, Zeb2/ZEB2) elevated by P. agglomerans. Cathelicidin's beneficial impact on the expression of genes/proteins involved in EMT was observed during and after the HP development; however, cathelicidin was not able to completely neutralize the negative changes. Nevertheless, significant EMT silencing in response to cathelicidin suggested the possibility of its use in the prevention/treatment of pulmonary fibrosis.

Pantoea agglomerans chronic exposure induces epithelial-mesenchymal transition in human lung epithelial cells and mice lungs.

Pantoea agglomerans is gram-negative bacteria widely distributed in nature. It predominates in inhalable dust from grain, herbs, and flax, and was identified as the most important cause of hypersensitivity pneumonitis (HP) in eastern Poland. To better understand the molecular mechanism of HP development studies focused on the interactions between P. agglomerans and alveolar epithelial cells as well as lung tissue with particular emphasis on the epithelial-mesenchymal transition (EMT). The studies were conducted on human normal lung epithelial NL20 cells and mice strain C57BL/6J. Cells and mice underwent chronic exposure to saline extract of P. agglomerans (SE-PA). Morphological changes were evaluated under light microscopy, the concentration of fibrosis markers was examined by the ELISA method, while the expression of genes involved in EMT was evaluated by RealTime PCR. During incubation with SE-PA epithelial cells underwent conversion and assumed fibroblast phenotype characterized by a decrease in epithelial cells markers (CDH1, CLDN1, JUP) and increase in mesenchymal cells markers (FN1, VIM, CDH2). Mice lungs collected after 14 days of SE-PA treatment revealed inflammation with marked lymphocytes infiltration. The intensified inflammatory process accompanied by increased proliferation of fibrous connective tissue was noted in mice lungs after 28 days of SE-PA exposure. Histological changes correlated with an increase of fibrosis markers (hydroxyproline, collagens), downregulation of epithelial markers (Cdh1, Cldn1, Jup, Ocln) and upregulation of myofibroblasts markers (Acta2, Cdh2, Fn1, Vim). Obtained results revealed SE-PA ability to induce EMT in human lung epithelial cells and mice lung tissue, with the scale of changes proportional to the time of treatment.

Neoadjuvant chemotherapy with or without oxaliplatin after short-course radiotherapy in high-risk rectal cancer: A subgroup analysis from a prospective study.

AIM: To evaluate the role of oxaliplatin in neoadjuvant chemotherapy delivered after short-course irradiation. BACKGROUND: Using oxaliplatin in the above setting is uncertain. PATIENTS AND METHODS: A subgroup of 136 patients managed by short-course radiotherapy and 3 cycles of consolidation chemotherapy within the framework of a randomised study was included in this post-hoc analysis. Sixty-seven patients received FOLFOX4 (oxaliplatin group) while oxaliplatin was omitted in the second period of accrual in 69 patients because of protocol amendment (fluorouracil-only group). RESULTS: Grade 3+ acute toxicity from neoadjuvant treatment was observed in 30% of patients in the oxaliplatin group vs. 16% in the fluorouracil-only group (p = 0.053). The corresponding proportions of patients having radical surgery or achieving complete pathological response were 72% vs. 77% (odds ratio [OR] = 0.88; 95% confidence interval [CI]: 0.39-1.98; p = 0.75) and 15% vs. 7% (OR = 2.25; 95% CI: 0.83-6.94; p = 0.16), respectively. The long-term outcomes were similar in the two groups. Overall and disease-free survival rates at 5 years were 63% vs. 56% (p = 0.78) and 49% vs. 44% (p = 0.59), respectively. The corresponding numbers for cumulative incidence of local failure or distant metastases were 33% vs. 38% (hazard ratio [HR] = 0.89; 95% CI: 0.52-1.52; p = 0.68) and 33% vs. 33% (HR = 0.78; 95% CI: 0.43-1.40; p = 0.41), respectively. CONCLUSION: Our findings do not support adding oxaliplatin to three cycles of chemotherapy delivered after short-course irradiation.

Drugs as important factors causing allergies.

Medications can cause many adverse reactions, both non-immunologic and immunologic ones. Allergies can take many forms, allergic reactions include all types of reactions according to Gell and Coombs. Typically, allergic reactions to drugs are manifested by skin lesions such as maculopapular rash or urticaria and life-threatening systemic reactions such as anaphylaxis. Allergy to drugs is diagnosed based on medical history and a number of specific tests: skin tests, blood tests. In diagnosing the causes of anaphylaxis, the basophil activation test is used to exclude false negative and false positive results of skin tests and specific IgE levels. Allergic reactions to medications usually resolve themselves after discontinuation of the drug. Sometimes in the treatment anti-allergic drugs are used to inhibit the development of skin lesions. After observing any signs of drug allergy it is important to accurately diagnose the cause, since the subsequent exposure to the drug may lead to a strong anaphylactic reaction and consequently death.

Age influence on hypersensitivity pneumonitis induced in mice by exposure to Pantoea agglomerans.

Hypersensitivity pneumonitis (HP) represents the immunologically mediated lung disease induced by repeated inhalations of a wide variety of certain finely dispersed organic antigens. In susceptible subjects, these inhalations provoke a hypersensitivity reaction characterized by intense inflammation of the terminal bronchioles, the interstitium and the alveolar tree. The inflammation often organizes into granulomas and may progress to pulmonary fibrosis. Our previous work indicated that cell extract of gram-negative bacteria Pantoea agglomerans (SE-PA) causes, in young C57BL/6J mice, pulmonary changes that are very similar to the clinical manifestations of HP in men. The purpose of presented studies was to describe the response of mice immune system while exposed to SE-PA. Particular attention was paid to examine the age influence on SE-PA induced inflammation and fibrosis in lung tissue. We used 3- and 18-month-old C57BL/6J mice. Lung samples were collected from untreated mice and animals exposed to harmful agent for 7 and 28 days. HP development was monitored by histological and biochemical evaluation. Using ELISA tests, we examined concentration of pro- and anti-inflammatory cytokines in lung homogenates. Our study demonstrated again that SE-PA provokes in mice changes typical for the clinical picture of HP, and that successive stages of disease (acute, subacute and chronic) might be obtained by modulation of time exposure. Furthermore, we found that animals' age at the time of sensitization influences the nature of observed changes (cytokine expression pattern) and the final outcome (reaction intensity and scale of fibrosis).

Farmers' occupational diseases of allergenic and zoonotic origin.

Farmers are exposed to a number of biological, physical and chemical factors harmful to the health, resulting from the specific nature of their work environment. As is clear from epidemiological studies about occupational diseases, pneumoconioses and infectious and parasitic diseases are recorded most commonly. Due to the character of farmer's work, the greatest risk to his health is biological hazards in the form of numerous microorganisms and their metabolites, and particles of plants and animals contained in the organic dust and pathogens transmitted by vectors such as ticks. The health status of farmers is often worse than of other professional groups as a result of limited access to health care and lower health literacy. Therefore, farmers should be subject to a system of diagnosing occupational diseases, and many preventive and educational programs concerning health risks associated with their work. The aim of this paper is to characterize occupational diseases of farmers including allergic diseases, tick-borne diseases and zoonoses.

Grain dust originating from organic and conventional farming as a potential source of biological agents causing respiratory diseases in farmers.

INTRODUCTION: Agricultural producers are exposed to a number of different health risks associated with their work environment. AIM: The objective of the study was to assess the degree of colonization by fungi in terms of quantity and in terms of variety of species the samples taken from the settled dust from combine threshing of rye cultivation from organic and conventional farms in the Province of Lublin. MATERIAL AND METHODS: This paper is a preliminary quantitative assessment of the species of fungi colonizing the samples of settled dust collected during combine threshing from organic and conventional farms in the Province of Lublin. One of the stages of the project was the classification of biosafety BSL (biosafety level) of selected isolates and API ZYM tests to evaluate the potential ability of isolates to cause adverse health effects. To determine the concentration and composition of fungi in collected samples plate dilution method was used with two media: Malt Agar and Potato Dextrose Agar. RESULTS: MOST COMMONLY ISOLATED FUNGI IN SETTLED DUST SAMPLES COLLECTED DURING COMBINE THRESHING FROM ORGANIC FARMS, ON PDA MEDIUM WERE: Alternaria alternata and Aureobasidium pullulans. Cultures on MA medium were dominated by Alternaria alternata, Mycelia sterilia and Fusarium poae. In samples of dust from conventional crops, the predominant species was Alternaria alternata on PDA medium and on MA medium. CONCLUSIONS: The obtained results show a potential risk of people involved in agricultural work.

Rye grains and the soil derived from under the organic and conventional rye crops as a potential source of biological agents causing respiratory diseases in farmers.

INTRODUCTION: Due to the specific work environment, farmers are exposed to various biological occupational hazard. Among these factors significant are fungi present in the grain and also in the soil. The fungi may be the cause of human diseases including skin infections, asthma, allergic rhinitis and many others. AIM: The aim of this study was to quantify and identify species of fungi colonizing rye grain samples and the soil under cultivation. MATERIAL AND METHODS: THE MATERIAL CONSISTED OF GRAIN AND SOIL SAMPLES FROM TWO AGRICULTURAL SYSTEMS: organic and conventional. To determine the concentration and composition of fungi in collected samples, two media: Malt Agar (MA, Becton, Dickinson and Company) and Potato Dextrose Agar (PDA, Becton, Dickinson and Company) were used. The composition of species in fungal flora was determined using macroscopic and microscopic methods. The isolates of fungi were ranked in the appropriate classes of biosafety BSL. RESULTS: THE MOST FREQUENTLY ISOLATED FUNGI FROM ORGANIC RYE GRAIN, REGARDLESS OF THE MEDIA USED, WERE SPECIES: Aureobasidium pullulans and Alternaria alternata. In conventional farms, most species isolated from rye grain were: Aureobasidium pullulans, Cladosporium oxysporum, Alternaria alternata and yeast-like fungi. Most often species isolated from the soil was Penicillium citreo-viride. CONCLUSIONS: All the results of the research demonstrate the potential hazard to the health of people working in agriculture. Significant exposure of this professional group is associated with the presence of harmful biological agents present in the grain and soil from its cultivation.

Allergy to selected cosmetic ingredients.

In an era in which cosmetics are commonly used, their often prolonged contact with the human body should determine the safety of their use. Often cosmetics are the cause of many side effects, mainly hypersensitivity reactions. Common groups of cosmetic components responsible for side effects are fragrances, preservatives and dyes. This paper focuses on the most allergenic components.

Beneficial impact of cathelicidin on hypersensitivity pneumonitis treatment-In vivo studies.

Cathelicidin (CRAMP) is a defence peptide with a wide range of biological responses including antimicrobial, immunomodulatory and wound healing. Due to its original properties the usefulness of CRAMP in the treatment of pulmonary fibrosis was assessed in a murine model of hypersensitivity pneumonitis (HP). The studies were conducted on mouse strain C57BL/6J exposed to a saline extract of Pantoea agglomerans cells (HP inducer). Cathelicidin was administered in the form of an aerosol during and after HP development. Changes in the composition of immune cell populations (NK cells, macrophages, lymphocytes: Tc, Th, Treg, B), were monitored in lung tissue by flow cytometry. Extracellular matrix deposition (collagens, hydroxyproline), the concentration of cytokines involved in inflammatory and the fibrosis process (IFNγ, TNFα, TGFß1, IL1ß, IL4, IL5, IL10, IL12α, IL13) were examined in lung homogenates by the ELISA method. Alterations in lung tissue morphology were examined in mouse lung sections stained with haematoxylin and eosin as well as Masson trichrome dyes. The performed studies revealed that cathelicidin did not cause any negative changes in lung morphology/structure, immune cell composition or cytokines production. At the same time, CRAMP attenuated the immune reaction induced by mice chronic exposure to P. agglomerans and inhibited hydroxyproline and collagen deposition in the lung tissue of mice treated with bacteria extract. The beneficial effect of CRAMP on HP treatment was associated with restoring the balance in quantity of immune cells, cytokines production and synthesis of extracellular matrix components. The presented study suggests the usefulness of cathelicidin in preventing lung fibrosis; however, cathelicidin was not able to reverse pathological changes completely.

Potential sources of infection with selected zoonotic agents in the veterinary work environment - pilot studies.

INTRODUCTION AND OBJECTIVE: The problem of occupational biohazards is very important, especially in the field of agriculture and in human and veterinary medicine. The aim of the study was to determine the potential sources of infection in veterinary professionals with selected zoonotic agents, including: Toxoplasma gondii, Giardia duodenalis, Leptospira spp., Cryptosporidium spp. and Coxiella burnetii. MATERIAL AND METHODS: A total of 50 air samples from barns, piggeries and veterinary surgeries were examined for the presence of Leptospira spp. and C. burnetii DNA. Serum samples of 86 pigs and 80 cows were tested for the presence of antibodies to Leptospira spp. and to phase I and II C. burnetii antigens. Serum of 70 cats were tested for the presence of antibodies to T. gondii and 65 samples of cat faeces for the presence of T. gondii oocysts. The presence of G. duodenalis and Cryptosporidium spp. were examined in 50 of dog faeces and 50 of bovine faeces samples. RESULTS: DNA of Leptospira spp. was detected in 2 air samples from the piggeries (4%). C. burnetii DNA was not found in any sample. Anti-Leptospira spp. antibodies were detected in 51 (59.3%) of examined pigs. Neither anti-Leptospira spp. nor anti-C. burnetii antibodies were found among samples of bovine serum. Anti-T. gondii antibodies was found in 52 cat serum samples (74.3%). Among samples of cat faeces, no T. gondii oocysts were detected. In one sample of cattle stool (2%), G. duodenalis was detected and in another (2%) - Cryptosporidium spp. G. duodenalis was detected in 7 samples (14%) and Cryptosporidium spp. in 2 samples (2%) of dog faeces. CONCLUSIONS: The results of this study demonstrate the potential risk of infection with Leptospira spp. in veterinarians working with pigs. Veterinarians could be also be at risk of infection with T. gondii and G. duodenalis.

Preliminary study on the occurrence of Toxoplasma gondii in Ixodes ricinus ticks from north-western Poland with the use of PCR.

A batch of 259 Ixodes ricinus ticks collected by flagging in woodlands of north-western Poland (Szczecin area) was examined for the presence of Toxoplasma gondii DNA with nested polymerase chain reaction (PCR). Positive isolates were genotyped with the restriction fragment length polymorphism (RFLP) analysis of B1 gene marker. Another batch of 340 I. ricinus ticks from Szczecin area was homogenized and inoculated into mice. Subsequently, the biological material obtained from inoculated mice was also examined with nested PCR for the presence of Toxoplasma gondii DNA and positive isolates were genotyped with RFLP as above. The study revealed that 12.7 % of I. ricinus ticks were infected with T. gondii. Prevalence of T. gondii infection was highest in females (27.9 %), being significantly greater (p<0.05) than in males (12.3 % ) and nymphs (5.6 % ). Most of the positive isolates (78.8 % ) belonged to type I of T. gondii. Samples from brains and other organs of 60 inoculated mice showed 44 positive results for T. gondii DNA with the dominance of atypical genotype and frequent mixed infections.

Study on Lyme borreliosis focus in the Lublin region (eastern Poland).

A suburban focus of Lyme borreliosis situated 11 km from the southern border of the city of Lublin (eastern Poland) was characterized. The focus covers an area of circa 100 km(2), surrounding 3 localities inhabited by circa 7,500 people engaged mostly in farming. It was demonstrated that on the area of focus the infection rate of Ixodes ricinus ticks with Borrelia burgdorferi, frequency of serological response of inhabitants to the antigen of Borrelia burgdorferi, and incidence of Lyme borreliosis were significantly (p<0.001) greater compared to the whole territory of Lublin province, and were respectively 13.1 % vs. 4.7 %, 33.0 % vs. 13.7 %, and 0.002 % vs. 0.00075 % .

Study on Giardia duodenalis and Cryptosporidium spp. infection in veterinarians in Poland.

The risk of exposure to zoonotic factors among veterinarians comprises still underestimated problem. Many etiological factors of infectious diseases are so far poorly known, including the way of their transmission from environment to humans and their impact for health. The main aim of the study was to determine the risk of two selected zoonosis infections caused by Giardia duodenalis and Cryptosporidium spp. among occupational group of veterinarians in Poland. Two hundred ninety seven samples of stool were tested for the presence of Giardia cysts and Cryptosporidium oocysts using Direct Fluorescent Assay (DFA). There were no positive results for Cryptosporidium. The presence of Giardia cysts was found in two samples of faeces (0.67%), confirmed by polymerase chain reaction and sequencing. The risk with regard to the parasites Giardia duodenalis and Cryptosporidium spp. seems to be low among the group of veterinarians.

Study on Toxoplasma Gondii, Leptospira Spp., Coxiella Burnetii, and Echinococcus Granulosus Infection in Veterinarians from Poland.

INTRODUCTION: Exposure to zoonotic factors in veterinary practice is closely related to the nature of the work. The main aim of the study was to determine the risk of selected zoonotic infections among the occupational group of veterinarians in Poland. MATERIAL AND METHODS: Blood samples of 373 veterinarians (162 males and 211 females) from 12 provinces of Poland were collected by the venipuncture of a forearm for serological tests. Commercial immunoenzymatic tests (ELISA) were used for detection of specific IgG antibodies to Echinococcus granulosus, IgM and IgG to Leptospira spp., and IgM, IgA, and I and II phase IgG to Coxiella burnetii. Enzyme-linked fluorescence assays (ELFA) were used to detect IgM and IgG antibodies to Toxoplasma gondii. RESULTS: Positive results were found in 209 (56.0%) veterinarians for at least one of the examined diseases. The overall proportion of participants found to have specific Toxoplasma gondii antibodies in the IgM and/or IgG assays amounted to 44.5%. The presence of Coxiella burnetii antibodies was found in 16 (4.3%) subjects, while Leptospira spp. antibodies were detected in 63 (16.9%) veterinarians. Among the 373 veterinarians examined, no Echinococcus granulosus antibodies were found. CONCLUSION: Results of the study seem to indicate a slightly elevated risk of Toxoplasma gondii infection and a moderate risk of infection with Leptospira spp. and Coxiella burnetii in veterinarians.

Prevalence of Anaplasma phagocytophilum in Ixodes ricinus ticks determined by polymerase chain reaction with two pairs of primers detecting 16S rRNA and ankA genes.

A total of 684 Ixodes ricinus ticks (321 nymphs, 184 males, and 179 females) were collected by flagging lower vegetation in 6 forest districts located on the territory of Lublin province (eastern Poland). Ticks were examined by polymerase chain reaction (PCR) method for the presence of Anaplasma phagocytophilum DNA with two pairs of primers: EHR521/EHR747 for detecting 16S rRNA gene, and LA6/LA1 for detecting ankA gene. To study the relationship between infection in ticks and people occupationally exposed to tick bite, blood serum samples of 261 forestry workers employed in the same forest districts were examined by immunofluorescence method for the presence of specific antibodies against A. phagocytophilum. A total of 70 ticks out of 684 examined (10.2%) showed the presence of A. phagocytophilum 16S rRNA gene. The prevalence of infection was significantly dependent on tick's stage (chi-square=49.2, p<0.00001) and geographical locality (chi-square=34.4, p<0.00001). The percentage of I. ricinus females infected with A. phagocytophilum (24.6%) was significantly greater compared to males (6.5%) and nymphs (4.4%) (p<0.00001). Only 19 ticks out of 684 examined (2.8%) showed the presence of A. phagocytophilum ankA gene, significantly less compared to 16S rRNA gene (p<0.00001). The prevalence of infection demonstrated by the presence of ankA gene was also significantly dependent on tick's stage (chi-square=23.6, p<0.00001) but not on locality (chi-square=9.8, p=0.082). A significant correlation was found between the presence of A. phagocytophilum 16S rRNA gene in I. ricinus female ticks from the particular forest districts and the serologic response to A. phagocytophilum of forestry workers employed in the same districts (p<0.05). No significant correlation was found between the presence of A. phagocytophilum ankA gene in I. ricinus ticks and serologic response of exposed workers. In conclusion, detection of A. phagocytophilum infection in ticks by PCR with the use of EHR521/EHR747 primers detecting 16S rRNA gene is significantly more sensitive compared to LA6/LA1 primers detecting ankA gene.

Prevalence of Borrelia burgdorferi genospecies in Ixodes ricinus ticks from Lublin region (eastern Poland).

The objective of the study was to determine the prevalence rate of 3 Borrelia burgdorferi genospecies in Ixodes ricinus ticks collected from wooded areas of the Lublin region (eastern Poland). A group of 1,813 I. ricinus ticks from 6 districts were examined for the presence of Borrelia burgdorferi sensu lato (B.b.s.l.) by polymerase chain reaction (PCR). Another group of 438 I. ricinus ticks collected from 4 districts were examined for the presence of B.b.s.l. by culture on BSKH liquid medium confirmed by PCR, and for the presence of Borrelia spp. by dark field microscopy (DFM). Borrelia burgdorferi genospecies (Borrelia burgdorferi sensu stricto, Borrelia afzelii and Borrelia garinii) were determined by nested-PCR in 113 ticks lysates showing presence of B.b.s.l. (in PCR or in culture and PCR). 5.4 % of I. ricinus ticks examined by PCR showed the presence of B.b.s.l. DNA. The infection rate was highest in females (12.1 %), lower in males (6.0 %) and the lowest in nymphs (1.7 %) (p < 0.001). The minimum infection rate of I. ricinus ticks with B.b.s.l. determined by culture was 3.4 %, whereas the minimum infection rate of ticks with motile spirochetes morphologically resembling Borrelia spp., determined by DFM, amounted to 11.2 %. The presence of all 3 Borrelia burgdorferi genospecies under investigation was found in ticks collected from 5 out of 6 examined districts. In 81.4 % of infected ticks only single infection with 1 genospecies was observed, while coinfections with 2 or 3 genospecies were detected respectively in 16.8 % and 1.8 % of infected ticks. Borrelia burgdorferi sensu stricto was the dominant genospecies in all examined tick stages and districts, both in single infections and in coinfections, and found in a total of 62.8 % of I. ricinus ticks infected with B.b.s.l. Borrelia afzelii and Borrelia garinii were less frequent and observed in respectively 39.8 % and 17.8 % of infected ticks.

Correlation of tests for detection of Borrelia burgdorferi sensu lato infection in patients with diagnosed borreliosis.

A group of 180 patients with diagnosed Lyme borreliosis were examined for the presence of infection with Borrelia burgdorferi sensu lato (B. burgdorferi s.l.) by serologic tests with B. burgdorferi s.l. antigens (IgM-ELISA, IgG-ELISA, IgM-immunoblot, IgG-immunoblot) and by polymerase chain reaction (PCR, nested-PCR) for detection of B. burgdorferi s.l. DNA in peripheral blood. A total of 61.7 %, 53.9 %, 62.2 %, and 59.4 % of the examined patients' sera showed positive or borderline results in the serologic tests IgM-ELISA, IgG-ELISA, IgM-immunoblot, and IgG immunoblot, respectively. The results of the tests IgM-ELISA and IgM-immunoblot were significantly correlated (p < 0.001). A higher degree of the correlation (p < 0.000001) was found at the comparison of results obtained with IgG-ELISA and IgG-immunoblot. The correlation between the positive findings in the IgM-ELISA and detection with IgM-immunoblot the diagnostically important B. burgdorferi s.l. OspC surface protein was relatively low but statistically significant (0.01 < p < 0.05). Much higher correlation was found between the positive findings in the IgG-ELISA and detection with IgG-immunoblot other diagnostically important B. burgdorferi s.l. antigen, the VlsE protein (p < 0.000001). The presence of B. burgdorferi s.l. DNA was found by PCR in 20 out 180 examined blood samples (11.1 %). No correlation was found to exist between the PCR results and the results of any of the serologic tests for detection of anti B. burgdorferi s.l. antibodies of IgM class. PCR results correlated significantly at a relatively low level (0.01 < p < 0.05) with the results of IgG-ELISA, but not with the results of IgG-immunoblot with regard to total reactions (0.2 < p < 0.1). By contrast, a distinctly significant correlation was found between the PCR results and detection of the VlsE protein with IgG-immunoblot (0.001 < p < 0.01). In conclusion, the results of the present study suggest that antibodies of IgG class are the most reliable marker in laboratory diagnostics of Lyme borreliosis, in particular those directed against VlsE surface protein of Borrelia burgdorferi sensu lato.

Prevalence of Babesia microti in Ixodes ricinus ticks from Lublin region (eastern Poland).

A total of 1,367 Ixodes ricinus ticks collected from 5 districts of the Lublin region (eastern Poland) were examined for the presence of Babesia microti DNA by PCR and nested-PCR. As many as 74 ticks (5.4 %) were found to be infected with Babesia microti. The infection rate varied significantly with stage/sex of ticks (chi (2) =16.48543, df=2, p < 0.000264). The infection rates in females and males amounted to 6.4 % and 8.8 % respectively and were significantly higher (p=0.006 and p=0.0001 respectively) compared to minimum infection rate in nymphs that was equal to 2.8 %. The prevalence of infection showed also a significant variability depending on geographic location within the Lublin region (chi(2) =18.62812, df=4, p < 0.000932). The highest rates of infection with Babesia microti were noted in ticks collected from the areas of Pulawy district situated in the northern part of region and the suburban Lublin district situated in the central part of the region (8.0 % and 7.3 % respectively). Mediocre infection rates (respectively 3.4 % and 3.3 %) were found in ticks from the Parczew and Wlodawa districts situated in eastern part of the region and covered with humid lakeland forests. The lowest infection rate (0.5 %) was noted in ticks from the Zamosc district situated in southern part of the region. In conclusion, the infection rate of Ixodes ricinus ticks with Babesia microti found in this study is higher compared to the majority of data reported by Polish and other European authors, and indicates a potential risk of human infection during occupational or recreational exposure to tick bite.