Gelatin medium for preserving of Trichinella spp. for quality control in laboratories - estimation of larvae viability.

INTRODUCTION AND OBJECTIVE: Official food control laboratories ensure food safety using reliable, validated methods. Council Regulations (EC) No. 853/2004, 854/2004 and 882/2004 of the European Parliament established hygiene rules the production of food of animal origin, together with requirements for official controls. This leads to detailed requirements for Trichinella control set out in Commission Implementing Regulation (EU) 2015/1375 of 10 August 2015. These regulations require the laboratory to participate in proficiency testing (PT) to confirm their competence and improve the quality of testing, and require the PT Organizer to use methods for the preparation and preservation of parasite larvae in order to evaluate and improve detection. Traditional methods of preparing such larvae expose them to rapid degradation, making it necessary to simultaneously isolate the larvae and place them in meatballs to ensure quality. MATERIAL AND METHODS: We developed a technique for preserving of Trichinella spp. for quality control such as PT sample preparation. The procedure protects larvae against toxic oxygen activity and bacterial destruction via a gelatin barrier. To estimate the viability of larvae preserved by this method, gelatin capsules with 10 larvae of T. spiralis in each were stored (4-8 °C) during 45 days of an experiment. Samples were tested at 2 day intervals (3 samples each day of testing). RESULTS: In total, 75 samples were tested. Larvae remained alive up to 3 weeks. The number of living larvae diminished after 27 days through day 43, after which no living larvae were observed. CONCLUSIONS: The gelatin medium procedure facilitated easy, high-throughput sample preparation and supported 100% recovery for 3 weeks. The method allows fast, efficient and accurate PT sample preparation.

Molecular confirmation of the systemic toxoplasmosis in cat.

INTRODUCTION AND OBJECTIVE: Systemic toxoplasmosis with tissue-spread parasites occurring in intermediate hosts may also occur in immunocompromised cats (e.g., infected with FLV or FIV). To the best of our knowledge, no reports have been published on the detection and genotyping of T. gondii DNA in cats with extraintestinal toxoplasmosis in Poland. The article describes the case of the sudden death of 3 out of 4 cats in a cattery, and the detection and molecular characterization of T. gondii DNA detected in the tissues of one of the dead cats. MATERIAL AND METHODS: Samples of brain, lungs, heart, and liver of the cat that died suddenly were examined for the presence of T. gondii DNA (B1 gene) by nested PCR and real-time PCR. DNA positive samples were also genotyped at 12 genetic markers using multiplex multilocus nested PCR-RFLP (Mn-PCR-RFLP) and multilocus sequence typing (MLST). RESULTS: A total of 9 out of the 20 DNA samples were successfully amplified with nested and/or Real-time PCR. DNA from 3 out of 5 types of tested samples were genotyped (brain, heart and muscle). Mn-PCR-RFLP and MLST results revealed type II (and II/III at SAG1) alleles at almost all loci, except a clonal type I allele at the APICO locus. This profile corresponds to the ToxoDB#3 genotype, commonly identified amongst cats in Central Europe. CONCLUSIONS: To the best of our knowledge, this is the first study describing the genetic characteristics of T. gondii population determined in a cat in Poland. These data confirm the importance of this host as a reservoir for this pathogen, and demonstrate the genotypic variation of this parasite. Veterinarians should take into account that cats may develop disseminated toxoplasmosis, and that it is a systemic disease which may lead to the death of the cat, and to transmission of the pathogen to other domestic animals and to humans.

Parasitological contamination of arable soil in selected regions of Poland - preliminary study.

INTRODUCTION AND OBJECTIVE: The hygienic status of arable soils in most developed countries has been unknown. In the presented study, a preliminary investigation was undertaken to determine the contamination with eggs of parasitic nematodes in the soil of arable fields in Poland. The aim of the study was to determine whether such contamination is common enough to constitute a significant problem and what factors may influence it. MATERIAL AND METHODS: The study was conducted in 5 Polish provinces from autumn 2021 to spring 2022. The provinces differed significantly in terms of the area of agricultural land, agricultural suitability, type of soil, scale of cattle and pig breeding, production of manure and slurry, and the use of manures and organic fertilizers for fertilization. A total of 133 soil samples were collected. Parasitological examination of soil samples was carried out using the PN-Z-19006 method [1], with confirmed high sensitivity. RESULTS: Parasite eggs were found in a total of 67 samples, of which 56 samples contained eggs of roundworms of the genus Ascaris (an average of 3.29 eggs/100 g of soil), 23 contained eggs of whipworms (an average of 1.22 eggs/100 g), and 3 contained eggs of Toxocara (1 egg/100 g). CONCLUSIONS: Differences in the percentage of positive samples were found depending on the period in which the samples were taken. The percentage of positive samples collected in autumn (53.57%) was higher than the percentage of positive samples collected in spring (48.05%). Similarly, the average number of eggs of in positive samples collected in autumn (3.43 eggs/100 g) was higher than the average number of eggs in samples collected in spring (2.90 eggs/100 g). Differences in the percentage of positive samples were also found depending on the region of origin of the samples.

Filters of automobile air conditioning systems as in-car source of exposure to infections and toxic moulds.

The main component of an air conditioning system is air filters. Over time, the filters of an air conditioning system in cars can turn into sources of emission of microbiological hazards. The aim of this study was to quantitatively and qualitatively assess the presence of infectious and toxic fungi in the AC filters in passenger cars. The studied non-woven filters were removed from passenger cars during the "winter"/"summer" seasons. The taxonomic identification of the fungi isolated from the filters was performed using both the culture-based and molecular methods. RT-PCR was applied to assess the presence of gene fragments regulating aflatoxin biosynthesis in the isolates obtained from fungal cultures. The average fungal concentrations in the filter samples collected during the summer/winter season were 5.4 × 104 cfu/m2 and 2.4 × 104 cfu/m2, respectively. Most of the filter samples, collected in both the studied seasons, revealed the presence of Aspergillus species including A. fumigatus, A. niger, A. terreus and/or A. flavus. The recorded levels of fungal contamination of AC filters in passenger cars indicate the necessity for more frequent filter replacement in this type of vehicle. Occupational exposure to moulds and the resulting health problems that may be experienced by professional drivers should be properly recognised in order to undertake effective preventive measures.

Knowledge, protection behaviours and seroprevalence of Lyme borreliosis in inhabitants of Lublin Province, eastern Poland - evaluation of a prophylaxis programme.

INTRODUCTION AND OBJECTIVE: Lyme borreliosis (LB) is the most frequent tick-borne disease with 17,338 cases reported in Poland in 2022. Since research on a LB vaccine is still ongoing, the promotion of individual behaviours and limiting of tick exposure is one of the most effective ways to prevent LB. The aim of the study was to evaluate the effectiveness of the LB prevention programme by assessing the knowledge, practice behaviours, seroprevalence of LB and satisfaction among the population of the Lublin Province in eastern Poland. MATERIAL AND METHODS: The prevention programme was carried out among 2,920 participants who were asked about their exposure to ticks, history of LB and prevention behaviours. Awareness of knowledge was evaluated before and after training. Seroprevalence of LB was rated by ELISA and immunoblot assays. RESULTS: Over 73% of participants reported tick bites in their lifetime, without significant differences between rural and urban area inhabitants. More than 80% of individuals declared that they use protective measures (PPM), such as proper clothes and body checking; repellents were the least frequently used by participants. The diagnosis of LB but not tick bites in a lifetime influenced the more frequent use of PPM. Increase in knowledge was observed in 86% of participants after education, and the highest knowledge was noted among respondents with higher education. The seroprevalence of anti-B. burgdorferi antibodies was 37% and was higher among men than women (40% vs. 36%). CONCLUSIONS: The population of Lublin Province is highly exposed to tick bites and infection with B. burgdorferi. The high seroprevalence and increase in knowledge confirmed the effectiveness and need for preventive programmes. These results can be useful for optimizing and enhancing the effects of future prevention campaigns.

Comparison of the effectiveness of various parasitological methods in detecting nematode eggs in different types of soil.

INTRODUCTION AND OBJECTIVE: Natural fertilizers, sewage sludge, digestates, as well as organic fertilizers produced on their basis, can become a source of parasitological contamination of cultivated land. High concentration of invasive forms of parasites in the soil may pose a threat to human and animal health. Therefore, it is necessary to control the hygienic condition of fertilizers and fertilized soils with particular emphasis on parasites. The aim of the study was to compare the effectiveness of methods commonly used for parasitological examination of soil with own methods which were used to develop the standards. MATERIAL AND METHODS: The study was carried out using samples of sandy soil (SS), horticultural mix soil (HS) and peat-based substrate (PS). Each sample was spiked with 100 dyed Ascaris suum eggs and examined with the use of 6 methods: Vasilkova, Dada, Quinn, and 3 methods according to the Polish Standards (PN-19000, PN- 19005, PN-19006). For each variant, 8 repetitions were made. RESULTS: The largest number of A. suum eggs were found with PN-19006 (mean number of detected eggs was 21.25, 46.50, 23.00 for HS, SS, PS, respectively. Slightly lower results were obtained using PN-19005 - the mean number eggs was 21.25, 36.00, 16.75, respectively. On the other hand, the mean number of A. suum eggs found with the Dada method was about 2-3 times lower than with the PN-19006 - 15.75, 22.50, 6.50 for HS, SS, PS soil, respectively. Other methods were much less effective. CONCLUSIONS: PN-19006 method turned out to be the most effective in detecting A. suum eggs. This method can be used for parasitological examination of soils and can be the basis for developing a system of methods dedicated to testing different types of soils for the presence of nematode eggs.

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.

The Consumption of Raw Goat Milk Resulted in TBE in Patients in Poland, 2022 "Case Report".

The alimentary route is the second most important route of tick-borne encephalitis infection. In Poland, the last TBE case due to the consumption of unpasteurized milk or dairy products of infected animals was recorded in 2017 as the fourth documented outbreak of TBEV infection in the country. In this study, two patients infected with TBEV through consumption of unpasteurized goat's milk from one source are described from a cluster of eight cases. In August and September 2022, a 63- and 67-year-old woman were hospitalized at the Infectious Diseases Clinic of the Institute of Rural Health (Lublin, Poland). The patients denied been recently bitten by a tick, and neither had been vaccinated against TBEV. The disease had a biphasic course. In the first case, the patient suffered from a fever, spine pain, and muscle weakness and paresis of the lower left limb. The second patient suffered from fever, vertigo, headaches, abdominal pain, and diarrhoea. The results of IgM and IgG antibodies were positive in both cases. After three weeks hospitalization, the patients were discharged in good condition. In one case, slight hearing impairment was observed. Vaccination and avoiding the consumption of unpasteurized milk remain the most effective ways to prevent tick-borne encephalitis.

Chlamydia psittaci in Faecal Samples of Feral Pigeons (Columba livia forma urbana) in Urban Areas of Lublin city, Poland.

Pigeons are a typical host and natural reservoir of Chlamydia psittaci, the etiological agent of avian chlamydiosis, considered as a neglected zoonotic diseases. The aim of the study was to determine the prevalence of C. psittaci in faecal samples of feral pigeons (Columba livia forma urbana) as a potential source of infection related to the presence of synanthropic birds in urban areas. A total of 143 samples of dry and fresh faeces of feral pigeons, were collected in the city of Lublin (Poland), from April to September 2021. Molecular detection of C. psittaci was performed by nested-PCR and real-time PCR, confirmed by sequencing. Among the collected samples, 5 positive results were obtained in nested-PCR (3.5%), while in real-time PCR, the number of positive samples increased to 11 (7.7%). The positive samples showed 100% identity to the C. psittaci strain AMK (CP047319.1). C. psittaci was found in 7 out of 111 (6.3%) faecal samples collected in public places, and in 4 out of 32 (12.5%) samples from the nesting site (4.9% and 2.8% among a total of 143 samples, respectively). The infection was detected in both dry and fresh faeces (9.1% and 4.5%, respectively). The highest number of positive results was obtained in June-5 (3.5%). Feral pigeons occurring in urban areas are a natural reservoir of C. psittaci posing a potential risk of zoonotic infections. However, further studies on exposure to contaminated pigeon faeces in terms of occupational and non-occupational risk of chlamydiosis are needed.

Mycobiota and mycotoxins in various kinds of vegetables and fruits as potential health risk factors for consumers - summary of a multiyear study.

INTRODUCTION: Between 2015-2020, the concentrations and diversity of filamentous fungi as well as the levels of mycotoxins in 4 classes of vegetables and fruits were studied in samples from farms located in eastern Poland. The short communication summarizes the results with the use of statistical analysis and indicates the potential health hazards associated with the consumption of the produce contaminated with fungi and/or mycotoxins. The concentrations of filamentous fungi in all examined samples were moderate (2.813 - 4.146 log10 CFU g -1). The highest values were noted in root vegetables, whereas the lowest values in fruit vegetables. The mycobiota revealed a marked biodiversity (67 species and 33 genera). Penicillium and Fusarium prevailed in the mycobiota of root vegetables, whereas Cladosporium and Alternaria in the remaining classes. Most of identified species (88.1%) were described as potentially pathogenic. The importance of mycotoxins as a potential health risk factor for vegetable consumers, was confirmed by the detection of a high prevalence (>55%) of total aflatoxin (AFT). The level of AFT in berry fruits was significantly higher than in other classes. The high prevalence (58.5%) of deoxynivalenol was noted in raspberries. CONCLUSIONS: Filamentous fungi present in the examined vegetables and fruits may pose a potential health risk for consumers due to a high prevalence of potentially pathogenic species, mostly those producing mycotoxins.

Mycobiota of berry fruits - levels of filamentous fungi and mycotoxins, composition of fungi, and analysis of potential health risk for consumers.

INTRODUCTION AND OBJECTIVE: The aim of the study was to determine the presence, concentration and generic composition of filamentous fungi. Selected mycotoxins were also determined: total aflatoxins and deoxynivalenol. MATERIAL AND METHODS: In 2017-2018, 40 samples of strawberry fruits and 40 samples of red raspberry fruits were collected. In 2019-2020, 37 samples of fresh strawberry fruits and 41 samples of fresh red raspberry fruits were collected on conventional farms located in eastern Poland and were subjected to mycological examination. The concentration and species composition of filamentous fungi were determined by the method of plate dilutions on malt agar. The isolated strains were identified using macroscopic and microscopic methods. Samples were also analysed for the presence of aflatoxin B1, total aflatoxin and deoxynivalenol using ELISA tests. RESULTS: The median concentrations of fungi were moderate or low: 3.079 and 3.491 log10 CFU g-1 for strawberries and raspberries, respectively. Fungi of the genus Cladosporium prevailed in the mycobiota of berry fruits, accounting for 84.3% of total isolates in raspberries and 34.8% in strawberries. The occurrence of total aflatoxins was detected in the majority of tested samples (73.2% in raspberries and 70.3% in strawberries) but never exceeded the level of 4.0 µg kg-1 assumed as safe. Deoxynivalenol has been detected only in raspberries with the prevalence of 58.5%. Its median concentration was 242.0 µg kg-1 and in 7 out of 41 samples (17.0%) exceeded the level of 750.0 µg kg-1, assumed as safe. CONCLUSIONS: Filamentous fungi and mycotoxins occurred in the examined berries at levels that mostly do not represent a health risk for immunocompetent people, but might pose such risk for immuno-compromised and/or atopic consumers.

Timber-colonizing gram-negative bacteria as potential causative agents of respiratory diseases in woodworkers.

OCCURRENCE: Gram-negative bacteria occur commonly in the inner tissues of stored coniferous and deciduous timber, showing a marked variation in numbers. The greatest maximal numbers are found in the sapwood of coniferous timber. The common constituents of the Gram-negative biota are potentially pathogenic species of Enterobacteriaceae family of the genera Rahnella, Pantoea, Enterobacter, and Klebsiella. The air of wood-processing facilities is polluted with the wood-borne Gram-negative bacteria and produced by them endotoxin, as demonstrated worldwide by numerous studies. EFFECTS: There are three potential pathways of the pathogenic impact of wood-borne Gram-negative bacteria on exposed woodworkers: allergic, immunotoxic, and infectious. Allergic impact has been underestimated for a long time with relation to Gram-negative bacteria. Hopefully, the recent demonstration of the first documented case of hypersensitivity pneumonitis (HP) in woodworkers caused by Pantoea agglomerans which developed in extremely large quantities in birch sapwood, would speed up finding of new wood-related cases of HP caused by Gram-negative bacteria. The second pathway is associated with endotoxin, exerting strong immunotoxic (excessively immunostimulative) action. It has been demonstrated that endotoxin is released into wood dust in the form of nano-sized microvesicles, by peeling off the outer membrane of bacteria. Endotoxin microvesicles are easily inhaled by humans together with dust because of small dimensions and aerodynamic shape. Afterwards, they cause a nonspecific activation of lung macrophages, which release numerous inflammatory mediators causing an inflammatory lung reaction, chest tightness, fever, gas exchange disorders, and bronchospasm, without radiographic changes. The resulting disease is known as "Organic Dust Toxic Syndrome" or "toxic pneumonitis." The potential third pathway of pathogenic impact is infection. The suspected species is Klebsiella pneumoniae that may occur commonly in wood dust; however, until now this pathway has not been confirmed. CONCLUSION: Summarizing, Gram-negative bacteria-inhabiting timber should be considered, besides filamentous fungi and actinobacteria, as important risk factors of occupational disease in woodworkers that could be either HP with allergenic background or toxic pneumonitis elicited by endotoxin.

Diversity and Drug Resistance of Filamentous Fungi Isolated from the Fresh Raspberries.

Fungi are one of the most widely distributed microorganisms in the environment, including food such as fruits, vegetables and other crops, posing a potential threat to food safety and human health. The aim of this study was to determine the diversity, intensity and drug resistance of potentially pathogenic filamentous fungi isolated from the fresh raspberries (Rubus idaeus L.). A total of 50 strains belonging to genera Fusarium, Cladosporium, Alternaria, Penicillium, Mucor, Rhizopus, Aspergillus and Acremonium were tested for drug resistance against 11 antifungals by disc diffusion and gradient strips methods. The average mycological contamination in the examined samples of raspberries amounted to 4.34 log CFU/g. The Cladosporium was isolated from all tested samples, followed by Alternaria and Fusarium with a frequency of 61% and 34%, respectively. The highest level of drug resistance was observed for Acremonium genera and Fusarium strains recorded a wide variation in drug resistance as revealed by susceptibility with amphotericin B and voriconzole with MICs ranged from 0.5-4 µg/ml and posaconazole with MICs ranging from 3-8 µg/ml. All fungal strains showed 100% resistance to caspofungin, fluconazole and flucytosine with both the methods, and 100% resistance to micafungin and anidulafungin in the gradient strip method.

Levels of filamentous fungi and selected mycotoxins in leafy and fruit vegetables and analysis of their potential health risk for consumers.

INTRODUCTION AND OBJECTIVE: The aim of the study was to determine the presence, concentration and generic composition of filamentous fungi. Considering the significant role of mycotoxins in the pathogenicity of fungal contaminants of vegetables, the scope of the study was extended by determination of aflatoxins and deoxynivalenol. MATERIAL AND METHODS: In the years 2019-2020, samples of vegetables (lettuce, spinach, tomato, red pepper) collected on conventional farms located in eastern Poland were subjected to mycological examination. The concentration and species composition of filamentous fungi were determined by the method of plate dilutions on malt agar. The isolated strains were identified with the use of macroscopic and microscopic methods. Samples were also analyzed for the presence of aflatoxin B1 (AFB1), total aflatoxin (AFT) and deoxynivalenol (DON) using the immunoenzymatic ELISA method. RESULTS: The median concentrations of filamentous fungi ranged from 2.778-3.204 log10 CFU g -1. Overall, 40 fungal species were identified in the examined vegetables, of which 38 are classified as potentially pathogenic for humans. The mean prevalence values for AFB1 and AFT were moderate or high (16.0-60.0% and 57.8-75.6%, respectively) and very low for DON (0-2.2%). The median concentrations of filamentous fungi, AFB1 and AFT were distinctly greater in leafy vegetables than on non-leafy tomato and pepper fruits, and the differences were highly significant (P<0.001). CONCLUSIONS: The levels of filamentous fungi and mycotoxins in Polish vegetables could be classified as moderate or low. The abundant presence of species with various pathogenic abilities may pose a risk for some categories of people consuming raw vegetables, mostly for immuno-compromised persons or atopics susceptible to food allergy caused by ingested moulds.

Is job seniority a protective factor against anxiety among midwives during the SARS-CoV-2 pandemic?

INTRODUCTION: Introduction. The pandemic SARS-Cov-2 outbreak necessitated the implementation of changes in everyday obstetric attendance, which demands continuity of care. Employment of sudden changes in obstetric attendance could have increased anxiety among medical personnel. OBJECTIVE: The aim of the Communication was to analyse the influence of professional experience on the level of anxiety in a group of midwives during the SARS-CoV-2 pandemic. MATERIAL AND METHODS: The analysis included 100 midwives: average professional experience: 11.51 years (max. 36; median 7,5; SD: 10.37; p<0.0001). The largest group of respondents was employed in a tertiary referral hospital (n = 40), the smallest in a medical clinic (n = 7). A preliminary study using an online questionnaire, the Generalized Anxiety Disorder Screener (GAD-7), posted on Internet fora, Facebook fan pages of The Childbirth with Dignity Foundation, and the Foundation for Midwives, using the Snowball sampling method. The Ethics Committee for Research Projects at the Institute of Psychology, University of Gdansk, approved the research project (Approval No. 35/2020). RESULTS: The average score was 9.390 (max. 21, median 8.5; SD; 5.228). The length of professional experience did not correspond to the level of anxiety (r = -0.0097; p = 0.9237). In 20 respondents, no anxiety was traced, while the smallest group indicated severe anxiety (n = 15). Professional experience did not influence the level of anxiety in either group with different level of anxiety (ANOVA test; p = 0.465). . CONCLUSIONS: Professional experience did not influence the level of anxiety. In order to prevent exacerbation of anxiety symptoms, different factors which may playa vital role in enhancing the level of anxiety must be analysed.

Microbial contamination of money sorting facilities.

INTRODUCTION: Money is the most common item with which we have daily contact. Circulated banknotes and coins can become microbiologically contaminated and act as both a source and a means of spreading such pollutants. MATERIAL AND METHODS: The study was carried out in three money sorting facilities in Poland. Bioaerosol samples were collected using a 6-stage Andersen impactor, and microorganisms deposited on tabletop surfaces were sampled using the swab method. Bacterial and fungal concentrations were calculated and all isolated species were taxonomically identified. RESULTS: The study confirmed that means of payment are active sources of microbial emission in money sorting facilities. The bioaerosol concentrations did not exceed the threshold limit values proposed for this type of office premises. It confirms that ventilation systems in these facilities worked efficiently, protecting them from the migration of microbial contaminants present in both indoor and outdoor (atmospheric) background air. On the other hand, the average concentrations of bacteria and fungi on tabletop surfaces in banknote and coin sorting rooms were above the proposed purity levels for indoor surfaces and should be treated as microbiologically contaminated. Microbiota isolated from the air and surfaces were very diverse and among those strains were bacterial and fungal pathogens that can pose a health threat to exposed individuals. CONCLUSIONS: The results showed that employees in money sorting facilities were exposed to microorganisms that may contribute to the development of adverse health outcomes. To protect them, highly efficient hygienic measures should be introduced in this working environment, to prevent both unwanted pollution and subsequent secondary emission of microbial contaminants from sorted means of payment and tabletop surfaces.

Toxoplasma gondii infection in slaughtered pigs and cattle in Poland: seroprevalence, molecular detection and characterization of parasites in meat.

BACKGROUND: Toxoplasma gondii infection may pose a severe medical problem especially in a congenital form and as an acquired infection in immunocompromised persons. Raw and undercooked meat of slaughtered animals is regarded as an important source of parasite infection; however, data concerning this issue in Poland are still insufficient. The aim of this study was to estimate the prevalence of T. gondii infection in pigs and cattle slaughtered for human consumption in Poland using serological and molecular methods. METHODS: Sera of 3111 pigs and 2411 cattle from 16 regions (voivodeships) of the country were examined for the presence of anti-T. gondii IgG using the direct agglutination test (DAT). Pepsin-digested samples of diaphragm and heart of seropositive animals were examined for the presence of T. gondii DNA (B1 gene) by nested PCR and real-time PCR, while non-digested samples were only examined by nested PCR. The B1 gene DNA samples were genotyped at 11 genetic markers using multilocus nested PCR-RFLP (Mn-PCR-RFLP) and sequencing. RESULTS: Seropositive DAT results were found in 11.9% of pigs and 13.0% of cattle. The highest seroprevalence was found in pigs from Podkarpackie (32.6%) and in cattle from Mazowieckie (44.6%). Data analysis showed that cattle > 5-10 years-old, as well as cattle and pigs from small farms, and pigs from farms with open production systems, had higher odds of testing seropositive (P < 0.05). Among the examined tissue samples, positive PCR results were found in samples from 12.2% and 10.2% of seropositive pigs and cattle, respectively. Among the samples successfully genotyped by Mn-PCR-RFLP and sequenced, four samples were identified as T. gondii type II and one sample as type I. CONCLUSIONS: The presence of T. gondii antibodies in a substantial proportion of examined pigs and cattle as well as the detection of parasite DNA in their tissues highlight a potential health risk to the consumers in Poland.

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.

Acute hypersensitivity pneumonitis in woodworkers caused by inhalation of birch dust contaminated with Pantoea agglomerans and Microbacterium barkeri.

CASE DESCRIPTION: Five workers (2 males and 3 females) employed in a furniture factory located in eastern Poland developed hypersensitivity pneumonitis (HP) after the pine wood used for furniture production was replaced by birch wood. All of them reported onset of respiratory and general symptoms (cough, shortness of breath, general malaise) after inhalation exposure to birch dust, showed crackles at auscultation, ground-glass attenuations in HRCT examination, and lymphocytosis in the BAL examination. The diagnosis of acute HP was set in 4 persons and the diagnosis of subacute HP in one. IDENTIFICATION OF SPECIFIC ALLERGEN: Samples of birch wood associated with evoking disease symptoms were subjected to microbiological analysis with the conventional and molecular methods. Two bacterial isolates were found to occur in large quantities (of the order 108 CFU/g) in examined samples: Gram-negative bacterium of the species Pantoea agglomerans and a non-filamentous Gram-positive actinobacterium of the species Microbacterium barkeri. In the test for inhibition of leukocyte migration, 4 out of 5 examined patients showed a positive reaction in the presence of P. agglomerans and 2 in the presence of M. barkeri. Only one person showed the presence of precipitins to P. agglomerans and none to M. barkeri. In the inhalation challenge, which is the most relevant allergological test in the HP diagnostics, all patients reacted positively to P. agglomerans and only one to M. barkeri. The results indicate that P. agglomerans developing in birch wood was the main agent causing HP in the workers exposed to the inhalation of dust from this wood, while the etiologic role of M. barkeri is probably secondary. CONCLUSION: The results demonstrate that apart from fungi and filamentous actinobacteria, regarded until recently as causative agents of HP in woodworkers, Gram-negative bacteria and non-filamentous actinobacteria may also elicit disease symptoms in the workers processing wood infected with large amounts of these microorganisms. The results obtained also seem to indicate that cellular-mediated reactions are more significant for causing disease symptoms compared to those that are precipitin-mediated.

Toxoplasma gondii infection in selected species of free-living animals in Poland.

INTRODUCTION AND OBJECTIVE: Free-living animals can play an important role as a reservoir of Toxoplasma gondi;, however, data concerning this issue in Poland are still limited.The aim of study was to assess the occurrence of T. gondii infection by using molecular methods in free-living animals in selected regions of Poland. MATERIAL AND METHODS: Tissues samples of 396 animals (foxes, muskrats, birds, martens, badgers, polecats, raccoons, minks, raccoon dogs, otters, small rodents and insectivores, and grass snakes were collected from various regions of Poland. After samples digestion, DNA was isolated using QIAmp DNA Mini Kit (Qiagen). DNA extraction from small rodents and insectivores samples was performed without digestion. Next, nested PCR (B1 gene) and, for a part of nested PCR positive amplicons, RFLP PCR, were performed according to the method by Grigg and Boothroyd (2001). The other part of nested PCR positive DNA isolates were genotyped using 5 genetic markers: SAG1, SAG2 (5'- and 3'), SAG3, BTUB and GRA6, based on the method by Dubey et al. (2006). These PCR products were sequenced and compared with the NCBI database using Blast. RESULTS: In total, in 50 of the 396 examined animals DNA of T. gondii was detected (12.6%). The highest percentages of positive results in PCR was obtained in martens (40.9%) and badgers (38.5%), lower in birds (27.3%) and the lowest in foxes (7.4%). The RFLP and multilocus PCR analysis showed the dominance of T. gondii clonal type II (or II/III). CONCLUSIONS: The results of this study indicate the frequent T. gondii infection among free-living animals in Poland, especially martens and badgers, which may indirectly indicate that these animals contribute to the spread of the parasite in the sylvatic environment in Poland. The genotyping analysis showed the dominance of T. gondii clonal type II (or II/III).