Comparison of the efficiency of two commercial kits – ELFA and Western blot in estimating the phase of <i>Toxoplasma gondii</i> infection in pregnant women

Sera of 89 pregnant women were selected according to the results of ELFA IgM, IgG and avidity IgG, and tested with commercial tests IgM, IgG and avidity IgG Western Blot (WB) to compare the efficacy of both techniques in determining the phase of T. gondii infection. In total, 81 of 89 tested sera (91.0%) were classified as positive, both in the ELFA and WB tests for the presence of anti- Toxoplasma antibodies of class IgG, indicating a past infection, while the prevalence of anti- Toxoplasma positive reactions associated with the antibodies of class IgM indicating a recent infection was much lower – 31.5% and 20.2%, respectively. Sera of 81 women were also tested in the ELFA and WB tests for avidity, e.g. ability of forming high-molecular IgG antibody complexes. Low or medium results in these tests (in this study all classified as low), indicating a recent infection, were detected by ELFA and WB in 22.2% and 45.7% of the total examined samples, respectively. The Spearman’s rank test for correlation, performed for recognition of quantitative data of the ELFA and WB tests (index, units or points), revealed a highly significant correlation between the ELFA and WB tests for homologous classes of antibodies, both for IgM and IgG (p0.05), except for the WB test for IgM antibodies, which showed a significant correlation with the ELFA test for IgG antibodies (p<0.01). A highly significant negative correlation between the ELFA and WB test for IgM antibodies and ELFA and WB tests for IgG avidity was demonstrated (p<0.01), except for a relationship between the WB test for IgM and WB for avidity, which was not significant. Such negative correlations are theoretically expected, as strong complexes with the participation of IgG antibodies are absent in the early phase of toxoplasmosis when early antibodies of IgM class are present. Summarizing, this study indicates the high usefulness of the commercial ELFA and WB tests in serodiagnostics of toxoplasmosis in pregnant women. Special attention should be paid to parallel detection of IgM antibodies and low values in the ELFA and WB tests for IgG avidity, which indicates a recent infection which may be associated with a clinical form of congenital toxoplasmosis and damage to the foetus

Identification of bacterial and fungal components in tobacco and tobacco smoke

The microbiological composition of tobacco products was studied using culture and chemical analysis (of tobacco leaves) or chemical analysis only (tobacco and tobacco smoke). The chemical analyses utilized gas chromatography-tandem mass spectrometry for determining 3-hydroxy fatty acids, muramic acid, and ergosterol as markers of respectively lipopolysaccharide (LPS), peptidoglycan, and fungal biomass. Mesophilic bacteria dominated in both fresh and cured tobacco leaves; a range of additional bacteria and fungi were also found albeit in minor amounts. The peptidoglycan and LPS concentrations were approximately the same in tobacco leaves as in cigarette tobacco. The concentrations of the measured microbial components were much lower in some cigarettes locally produced in China, Korea, and Vietnam than in cigarettes of international brands purchased in the same countries, and the concentrations in the smoke were in general agreement with the concentrations in cigarette tobacco. No differences in microbial load in tobacco of "light" and "full flavor" cigarettes were seen. Storing cigarettes at high humidity resulted in elevated levels of fungi in the cigarette tobacco leading to increased ergosterol concentrations in the smoke. The fact that tobacco smoke is a bioaerosol may help to explain the high prevalence of respiratory disorders among smokers and non-smokers exposed to second hand smoke since the same symptoms are also commonly associated with exposure to bioaerosols

Prevalence of tick-borne pathogens at various workplaces in forest exploitation environment

Background: The objective of the study was the evaluation of the infection of ticks with pathogenic microorganisms at various workplaces (timber acquisition, forest growing, forest cultivation, forest protection). Material and Methods: Eight hundred sixty one Ixodes ricinus ticks collected from 4 workplaces were examined for the presence of Borrelia burgdorferi, Anaplasma phagocytophilum and Babesia microti by polymerase chain reaction (PCR). Then, a comparative analysis of the relative density and infection of ticks at individual workplaces was done. In the statistical analysis, Chi2 test, and Pearson’s test for correlation were applied. Results: The differences in infection (15.9–50%) of ticks with B. burgdorferi between the examined workplaces were highly significant, with the highest percentage observed at forest growing. The percentages of infection of ticks with A. phagocytophilum at individual workplaces ranged from 1.1–3.7%, and differences were statistically insignificant. The percentages of infections of ticks with Babesia microti at individual workplaces fluctuated from 3.6–4.4% and differences were also insignificant. Co-infections of ticks with 2 or 3 pathogens were rare. Conclusions: Co-infections with B. burgdorferi and B. microti showed a significant relationship with the workplaces, while those with B. burgdorferi and A. phagocytophilum did not show such a dependence. No significant positive correlation was found between the relative density of ticks and the frequencies of infections with B. burgdorferi, A. phagocytophilum and B. microti. Med. Pr. 2014;65(5):575–58

Risk factors for work-related eczema and urticaria among vocational students of agriculture

Introduction and objective. Farmers are at high risk of occupational skin diseases which may start already during vocational training. This study was aimed at identification of risk factors for work-related skin diseases among vocational students of agriculture. Materials and method. The study involved 440 students (245 males, 195 females aged 17–21 years) in 11 vocational schools which were at least 100 km from each other. The protocol included a physician-managed questionnaire and medical examination, skin prick tests, patch tests, total IgE and Phadiatop. Logistic regression model was used for the identification of relevant risk factors. Results. Work-related dermatoses were diagnosed in 29 study participants (6.6%, 95%CI: 4.3–8.9%): eczema in 22, urticaria in 14, and co-existence of both in 7 students. Significant risk factors for work-related eczema were: history of respiratory allergy (OR=10.10; p<0.001), history of eczema (itchy rash) provoked by wet work and detergents before entering the school (OR=5.85; p<0.001), as well as history of contact dermatitis to metals, rubber or cosmetics prior to inscription (OR=2.84; p=0.016), and family history of any skin disease (OR=2.99; p=0.013). Significant risk factors for work-related urticaria were: history of allergic rhinitis and asthma prior to inscription (OR=7.29; p=0.006), positive skin prick tests to work place allergens (OR=4.65; p=0.002) and to environmental allergens (OR=3.79; p=0.009), and positive Phadiatop test (OR=3.61; p=0.013). Conclusions. Work-related skin diseases are common among vocational students of agriculture. Atopy, past history of asthma, allergic rhinitis, and eczema (either atopic, allergic or irritant) are relevant risk factors for work-related eczema and urticaria in young farmers, along with family history of any skin disease. Positive skin prick tests seem relevant, especially in the case of urticaria. Asking simple, aimed questions during health checks while enrolling students into agricultural schools would suffice to identify students at risk for work-related eczema and urticaria, giving them the chance for selecting a safer profession, and hopefully avoiding an occupational disease in the future

Microbiological characterization of vegetables and their rhizosphere soil in Eastern Poland

The aim of this study was to investigate the bacteriological quality of 5 kinds of vegetables (lettuce, dill, radish, beetroot, carrot) and their rhizosphere soil, originating from conventional farms located in the Lublin Province of Eastern Poland. A total number of 35 samples of fresh vegetables (FV) taken immediately from soil, 35 samples of soil from rhizosphere of these vegetables (SR) and 35 samples of vegetables sold at retail in the markets in Lublin (VR) were examined. The samples were analysed for the content of: aerobic mesophilic bacteria (AMB) grown at 30°C and 37°C, Gram-negative bacteria of Enterobacteriaceae family, faecal coliform (FC) bacteria, Salmonella spp., and Clostridium perfringens. Median AMB values determined at 30°C for FV, SR and VR were 5.27, 5.00, and 5.00 log 10 CFU g -1 , respectively, being significantly greater compared to those recorded at 37°C. The exceeding of the threshold value of 6.0 log 10 CFU g -1 proposed by Gelosa (1998) was noted only in 5 FV samples grown at 30°C (14.3%), and in 3 FV samples grown at 37°C (8.6%). The threshold value was never exceeded in SR and VR samples. Median concentrations of Enterobacteriaceae determined for FV, SR and VR were 4.03, 3.87, and 3.04 log 10 CFU g -1 , respectively. Eleven species of Enterobacteriaceae were identified in the FV, SR and VR samples. The percent of samples containing Escherichia coli was greatest for VR (22.9%), smaller for FV (17.1%) and smallest for SR (5.7%). The median concentrations of the faecal coliform bacteria (FC), determined by culture at 44°C, were low, amounting to 1.000 log 10 CFU g -1 for FV and SR and 0.00 for VR. All examined vegetable and soil samples tested negative for the presence of Salmonella. The median concentrations of Clostridium perfringens were low, amounting to 0.00 log 10 CFU g -1 for all categories of samples. This bacterium was relatively common in soil samples with the prevalence of 40.0%, but very rare in vegetable samples (occurring in 5.7% of FV and in none of VR samples). In conclusion, the results of the present study generally indicate that the microbiological quality of Polish vegetables grown on conventional farms is satisfactory and safe for consumers

Age influence on mice lung tissue response to <i>Aspergillus fumigatus</i> chronic exposure

[b]Introduction and objective[/b]. Exposure to conidia of [i]Aspergillus fumigatus[/i] was described as a causative factor of a number of the respiratory system diseases, including asthma, chronic eosinophilic pneumonia, hypersensitivity pneumonitis and bronchopulmonary aspergillosis. The study investigates the effects of the repeated exposure to [i]A. fumigatus[/i] in mice pulmonary compartment. Our work tackles two, so far insufficiently addressed, important aspects of interaction between affected organism and[i] A. fumigatus[/i]: 1) recurrent character of exposure (characteristic for pathomechanism of the abovementioned disease states) and 2) impact of aging, potentially important for the differentiation response to an antigen. [b]Materials and methods[/b]. In order to dissect alterations of the immune system involved with both aging and chronic exposure to [i]A. fumigatus[/i], we used 3- and 18-month-old C57BL/6J mice exposed to repeated[i] A. fumigatus[/i] inhalations for 7 and 28 days. Changes in lung tissue were monitored by histological and biochemical evaluation. Concentration of pro- and anti-inflammatory cytokines in lung homogenates was assessed by ELISA tests. [b]Results and conclusions. [/b]Our study demonstrated that chronic inflammation in pulmonary compartment, characterized by the significant increase of proinflammatory cytokines (IL1, IL6, IL10) levels, was the dominant feature of mice response to repeated [i]A. fumigatus[/i] inhalations. The pattern of cytokines' profile in the course of exposure was similar in both age groups, however in old mice the growth of the cytokines' levels was more pronounced (especially in case of IL1)

Prevalence of infections and co-infections with 6 pathogens in <i>Dermacentor reticulatus</i> ticks collected in eastern Poland

Occurrence of co-infections with various pathogens in ixodid ticks creates a risk of increased severity of tick-borne diseases in humans and animals exposed to bite of the ticks carrying multiple pathogens. Accordingly, co-infections in ticks were subject of numerous analyses, but almost exclusively with regard to Ixodes ricinus complex whereas potential tick vectors belonging to other genera were much less studied. Taking into consideration the role of Dermacentor reticulatus in the transmission of various pathogens, we carried out for the first time the comprehensive statistical analysis of co-infections occurring in this tick species. An attempt was made to determine the significance of the associations between 6 different pathogens occurring in D. reticulatus (Tick-borne encephalitis virus = TBEV, Anaplasma phagocytophilum , Rickettsia raoultii , Borrelia burgdorferi s. l., Babesia spp., Toxoplasma gondii ), using 2 statistical methods: determination of Odds Ratios (ORs) and the Fisher’s exact test. 634 questing Dermacentor reticulatus ticks (370 females and 264 males) were collected in 2011– 2013 by flagging the lower vegetation in 3 localities in the area of Łęczyńsko-Włodawskie Lakeland, situated in the Lublin region of eastern Poland. The presence of individual pathogens was detected by PCR. Ticks were infected most often with Rickettsia raoultii (43.8%), less with TBEV (8.5%), and much less with Babesia spp., Toxoplasma gondii , Borrelia burgdorferi s.l., and Anaplasma phagocytophilum (2.5%, 2.1%, 1.6% and 1.1%, respectively). The locality-dependent variability proved to be significant for TBEV (χ 2 =11.063; P=0.004) and Toxoplasma gondii (χ 2 =11.298; P=0.0035), but not for other pathogens. Two hundred seventy (42.6%) of the examined ticks were infected only with a single pathogen, and 54 (8.5%) showed the presence of dual co-infections, each with 2 pathogens. The most common were dual infections with participation of Rickettsia raoultii (7.41%); next, those with participation of the TBEV (5.21%), Toxoplasma gondii (1.58%), Borrelia burgdorferi s.l. (1.26%), Anaplasma phagocytophilum (0.95%), and Babesia spp. (0.63%). On the total number of 15 possible associations, in 9 cases co-infections occurred whereas in 6 cases they were not detected. The most noteworthy were positive co-infections with the participation of TBEV, which proved to be weakly significant (0.05&lt;P&lt;0.1) in associations with Toxoplasma gondii and Anaplasma phagocytophilum , with Odds Ratios over 3.3 and 4.4, respectively. The values of Odds Ratios exceeded 3.0 also at the co-infections of Rickettsia raoultii with B. burgdorferi s.l., and T. gondii with Babesia spp., but these associations did not attain a significance level. The co-infections of Rickettsia raoultii with Babesia spp. appeared not to be significant (0.05&lt;P&lt;0.1) with OR below 0.3. In conclusion, co-infections with various pathogens in D. reticulatus ticks seem to be relatively rare and mostly not significant

Defining a novel domain that provides an essential contribution to site-specific interaction of Rep protein with DNA

15 p.-6 fig.An essential feature of replication initiation proteins is their ability to bind to DNA. In this work, we describe a new domain that contributes to a replication initiator sequence-specific interaction with DNA. Applying biochemical assays and structure prediction methods coupled with DNA-protein crosslinking, mass spectrometry, and construction and analysis of mutant proteins, we identified that the replication initiator of the broad host range plasmid RK2, in addition to two winged helix domains, contains a third DNA-binding domain. The phylogenetic analysis revealed that the composition of this unique domain is typical within the described TrfA-like protein family. Both in vitro and in vivo experiments involving the constructed TrfA mutant proteins showed that the newly identified domain is essential for the formation of the protein complex with DNA, contributes to the avidity for interaction with DNA, and the replication activity of the initiator. The analysis of mutant proteins, each containing a single substitution, showed that each of the three domains composing TrfA is essential for the formation of the protein complex with DNA. Furthermore, the new domain, along with the winged helix domains, contributes to the sequence specificity of replication initiator interaction within the plasmid replication origin.National Science Centre [2012/04/A/NZ1/00048 to I.K.;2017/26/D/NZ1/00239 to K.W.]; Foundation for Polish Science [TEAM, POIR.04.04.00-00-5C75/17-00 to I.K.]; International Institute of Molecular and Cell Biology in Warsaw (to J.M.B.); Ministerio de Economía,Industria y Competitividad (MINECO/AEI) [BIO2012-30852, RTI2018-094549-B-I00 to R.G.]. Funding for open access charge: Foundation for Polish Science [TEAM,POIR.04.04.00-00-5C75/17-00].Peer reviewe