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.

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.

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.

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.

Contamination of soil with eggs of geohelminths Ascaris spp., Trichuris spp., Toxocara spp. in Poland - potential source of health risk in farmers.

Soil is one of the most commonly occurring sources of biological hazards in the environment. It is a reservoir of many pathogens and an important factor in the environmental transmission of parasites. The aim of the article is to discuss the general degree of parasitic contamination of agriculturally used soil in Poland from the aspect of potential health risk for farmers. Parasitic studies of soil conducted to-date in Poland demonstrate that it is considerably contaminated. The presence of the eggs of geohelminths Ascaris spp., Toxocara spp., and Trichuris spp was most often observed. The main cause of contamination of agriculturally used soil are fertilizers, sewage sludge, as well as animal faeces. The presence of invasive forms of parasites in the environment creates the risk of infection in humans due to direct or indirect exposure resulting from the way of performing work. In Poland, the frequency of infections with nematodes of the genera Ascaris spp., Trichuris spp., Toxocara spp. in humans has not been recognized. Considering the high percentage of positive results obtained in parasitic examinations of soil it may be presumed that there is still a real threat to human health, including population groups engaged in agricultural production.

Parasitological contamination with eggs Ascaris spp., Trichuris spp. and Toxocara spp. of dehydrated municipal sewage sludge in Poland.

The objective of the present study was to evaluate the contamination of sewage sludge produced by municipal waste treatment plants in Poland by viable eggs of intestinal parasites of the genera Ascaris, Toxocara and Trichuris (ATT). Ninety-two municipal, mechanical-biological sewage treatment plants located within Poland were selected. These plants belonged to types of agglomerations: group 0 (large), group 1 (medium), group 2 (smaller) and group 3 (small). Samples were collected at the final stage of sewage treatment after the addition of flocculent to sludge, followed by dehydration. The samples were examined by a method adjusted to examine sewage sludge dehydrated using polyelectrolytes. The viability of the isolated eggs was evaluated based on incubation in a moist chamber. Live eggs of intestinal nematodes were found in 99% of samples. Most samples were contaminated by the eggs of Ascaris spp. (95%) and Toxocara spp. (96%). However, Trichuris spp. eggs were detected in 60% of samples. The mean number of eggs in 1 kg of dry mass (eggs/kg d.m.) was 5600 for Ascaris, 3700 for Toxocara and 1100 for Trichuris. The highest number of ATT eggs was detected in samples from sewage treatment plants located in south-eastern and central Poland. The highest number of ATT eggs was found in sewage sludge produced in large sewage treatment plants (agglomeration Groups 0 and 1), with mean values of 15,000 and 8900 eggs/kg d.m. The present study is the first parasitological investigation conducted on a large number of samples (92 samples) taken from various types of municipal sewage treatment plants located throughout Poland (16 regions) after the common introduction of polyelectrolytes during sewage sludge dehydration. The results of this study indicate that sludge produced in municipal sewage treatment plants is highly contaminated with parasite eggs.

Negative effect of flocculant (cationic acrylamide) on detectability of the nematode eggs in sewage sludge.

The use of sewage sludge in agriculture brings the risk of microbiological and parasitological contamination of soil, ground and surface water, as well as cultivated plants. Therefore prior the application to the soil, sewage sludge must be examined, among others, for the presence of live eggs of intestinal parasites. However, the efficiency of commonly used for this purpose parasitological methods is not satisfactory. This is probably due to the presence of flocculants in the sediments used in the dehydration process. The objective of the study was analysis of the effect of flocculant (cationic acrylamide) on the possibilities of isolation of parasite eggs from dehydrated sewage sludge. For this purpose 10 samples of sewage sludge were prepared: 5 containing flocculant and 5 without flocculant. Samples were tested by flotation method according to Quinn. From sewage sludge free of flocculant, 67.8 eggs were isolated, on average, whereas from sludge containing flocculant - only 2.8 eggs. The experiments confirmed that the isolation of eggs from sewage sludge containing flocculant (cationic acrylamide) is much more difficult than from sludge free from this substance and therefore the simple parasitological methods should not be used to examine the dehydrated sewage sludge.

Contamination of selected recreational areas in Lublin Province, Eastern Poland, by eggs of Toxocara spp., Ancylostoma spp. and Trichuris spp.

INTRODUCTION: While using recreational areas, people take with them accompanying animals (dogs, cats). These animals are the main source and reservoir of dangerous zoonoses, including parasitoses caused by genera of nematodes, also called roundworms, Toxocara spp., Trichuris spp. and Ancylostoma spp. Attention should also be paid to the possibility of contamination of recreational areas with the eggs of intestinal parasites by wild animals (e.g. red fox). MATERIAL AND METHODS: Sand was collected on beaches and leisure terrains located in recreational areas in Zwierzyniec, Rudka reservoir on the river Wieprz, 'Echo' reservoir, accessible to tourists to the Roztocze National Park, and the 'Florianiecki' reservoir, chich is inaccessible to tourists due to its location in the depths of the National Park. Material for the research was also collected from the 'Zemborzycki reservoir in Lublin, A total of 300 sand samples were collected. The samples, approximately 500 g each, were collected into plastic bags from the superficial layer of sand, at a depth up to 3 cm. For egg detection, flotation with the Wasylikowa metod and the modified Quinn method were used. In the Quinn metod, the flotation solution is NaCl saturated solution, in the Wasylikowa method, 5% solution of NaOH is used. RESULTS: The most contaminated grounds were areas by the water 'Zemborzycki' reservoir in Lublin. The eggs of intestinal parasites were isolated from 13 out of 24 samples examined. In sand collected from the beach by the 'Echo', 'Florianiecki' and 'Rudka' water resevoirs in Zwierzyniec, no eggs of intestinal parasites were detected. CONCLUSIONS: Such a dissemination of results may indicate a connection between the degree of occurrence of geohelmint eggs and the intensity of human movement, together with accompanying animals, in places that can be used for recreation.

Fungal diversity of root vegetables and soil rhizosphere collected from organic and conventional farms in Eastern Poland.

OBJECTIVE: Determination of the concentration and species composition of filamentous fungi in root vegetables (carrots and red beetroots), and the adjacent rhizosphere soil, collected on organic and conventional farms in Eastern Poland. MATERIAL AND METHODS: During the period 2015-2016, a total number of 80 samples of vegetables and 40 samples of soil were examined. From each type of farm, 20 samples of vegetables and 20 samples of the adjacent soil were examined. In addition, the study included 20 samples of vegetables from organic farms and 20 samples of vegetables from conventional farms purchased on the markets in the city of Lublin in Eastern Poland. In order to determine the concentration and species composition of filamentous fungi, both in vegetables and soil, the method of plate dilutions on Malt Agar medium (Difco) with chloramphenicol was used. The study was conducted in two parallel repetitions. Inoculated media were incubated at the temperature of 30° C for 72 hours, then at room temperature for 72 hours. The species composition of fungal flora was determined using macroscopic and microscopic methods, with the help of keys and atlases. RESULTS: It was found that the mean concentration of fungi was higher in vegetables and soil from conventional farms than in those from organic farms. In the case of carrots and soil from conventional farms, this concentration was 4.93 and 5.10 log10 CFU g-1, respectively, whereas from organic farms - respectively, 3.81 and 4.20 log10 CFU g-1. In the case of beetroots and soil from conventional farms, the mean concentrations were also higher compared to organic farms - 5.09 vs. 3.93 and 4.95 vs. 4.23 log10 CFU g-1. In the examined vegetables and rhizosphere soil, 61 species of filamentous fungi were found, of which 12 belonged to the genus Penicillium, 4 to the genus Fusarium, and 2 species each to the genera: Aspergillus, Cladosporium and Trichoderma. As many as 28 (45.9%) fungal species that occurred in vegetables and soil are regarded as pathogenic for humans.

Variations in the Rate of Infestations of Dogs with Zoonotic Nematodes and the Contamination of Soil in Different Environments.

Companion animals are an important aspect in human life. However, they may also be considered a source of pathogens. An example of zoonotic parasitoses is toxocarosis or cutaneous larva migrans (CLM). The aim of the study was to detect zoonotic nematodes of dogs living in different areas and the intensity of contamination in parasite polluted environments that are hazardous to human health. The fecal samples were examined using standard flotation and decantation methods as well as McMaster's quantitative technique. The soil samples in urban and rural areas were examined using a modified flotation method as described by Quinn et al. Statistical analyses were performed by IBM SPSS Statistics Version 23. The overall prevalence of parasites in dogs was 38%, 17.02% and 56.60% from urban and rural areas, respectively. The percentage values of nematodes important for human health (Toxocaracanis, Ancylostomatidae, Trichurisvulpis) remained at the same level (16%). The infected dogs were dominated by a single parasite species, the main was T.canis (28.95%). In total, 54.30% of the soil samples were contaminated with parasite eggs. The contamination of urban and rural sandpits was 40% and 60%, respectively. The molecular examinations of soil samples using LAMP (loop-mediated isothermal amplification) confirmed the presence of nematode eggs of the species T.canis in all samples previously classified as positive.

Viability assessment of Ascaris suum eggs stained with fluorescent dyes using digital colorimetric analysis.

The aim of the study was to develop a method for the colorimetric evaluation of nematode eggs using appropriate instruments. The materials for the study were live and dead (inactivated) eggs of the Ascaris suum. Viability of the eggs was assessed using four different kits for fluorescent staining (for each technique, a series of photos were taken). Images of stained eggs were analysed using graphic software with RGB (red-green-blue) function. The viability of the eggs was assessed according to the relative positions of the distributions of colour intensities of live or dead eggs - distributions area's overlap index (DAOI), and distributions area's separation index (DASI) were calculated. Computer analysis of the intensity of green colour was not satisfactory. However, analysis of images in the spectrum of red colour proved useful for the effective differentiation between live or dead eggs. The best parameters were observed using the Annexin V FITC Apoptosis Detection Kit (DASI = 41 and 67). The investigation confirmed the usefulness of fluorescent dyes used in conjunction with digital analysis for the assessment of the viability of A. suum eggs. The use of computer software allowed a better objectivity of the assessment, especially in the case of doubtful staining.

Division of methods for counting helminths' eggs and the problem of efficiency of these methods.

From the sanitary and epidemiological aspects, information concerning the developmental forms of intestinal parasites, especially the eggs of helminths present in our environment in: water, soil, sandpits, sewage sludge, crops watered with wastewater are very important. The methods described in the relevant literature may be classified in various ways, primarily according to the methodology of the preparation of samples from environmental matrices prepared for analysis, and the sole methods of counting and chambers/instruments used for this purpose. In addition, there is a possibility to perform the classification of the research methods analyzed from the aspect of the method and time of identification of the individuals counted, or the necessity for staining them. Standard methods for identification of helminths' eggs from environmental matrices are usually characterized by low efficiency, i.e. from 30% to approximately 80%. The efficiency of the method applied may be measured in a dual way, either by using the method of internal standard or the 'Split/Spike' method. While measuring simultaneously in an examined object the efficiency of the method and the number of eggs, the 'actual' number of eggs may be calculated by multiplying the obtained value of the discovered eggs of helminths by inverse efficiency.

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 log10 CFU g-1 , respectively, being significantly greater compared to those recorded at 37°C. The exceeding of the threshold value of 6.0 log10 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 log10 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 log10 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 log10 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.

Characterisation of a new, highly effective method for detecting nematode eggs (Ascaris spp., Toxocara spp., Trichuris spp.) in sewage sludge containing flocculants.

Because traditional methods used for sewage sludge parasitological examinations have low sensitivity, a new, highly effective method (own method - OM) was devised. The principle of this method is to eliminate the flocculent effect on the structure of sewage sludge by mechanically damaging floccules in the presence of surfactants and to increase the effectiveness of egg isolation processes in large volumes of liquids. The objective of this study was to estimate the effectiveness of the OM in detecting nematode eggs in sewage sludge samples containing flocculants. In the first stage, the effectiveness of the OM was compared to 4 other methods routinely used in parasitological examinations of dehydrated sewage sludge. Next, method standardisation was performed using sewage sludge samples supplemented with eggs from 3 parasite species (Ascaris suum, Toxocara canis and Trichuris vulpis). The study demonstrated that OM efficiency was 6-65 times greater than other methods, depending on the method and type of detected eggs. Limit of detection (LOD) calculations for the OM were performed on samples supplemented with a known number of parasite eggs resulting in 10, 5 and 3 eggs/50 g of sample for A. suum, T. vulpis and T. canis eggs, respectively. The limits of quantification (LOQ) of the OM were established as 200 eggs/50 g of sample for A. suum and T. vulpis eggs and 50 eggs/50 g of sample for T. canis eggs. The rectilinear regression functions, which determined the relationship between the number of eggs detected in OM measurements and the number of eggs contained in the samples, were characterised by high and statistically significant coefficients of determination (r2). The slopes of the trend lines were 0.3188, 0.3821 and 0.3276, and the intercepts were -11.223, -9.0261 and -23.15 for A. suum, T. canis and T. vulpis eggs, respectively. Method sensitivity, calculated as the slope coefficient of the regression function and expressed as a percentage, ranged from 32% to 38% depending on egg type. The study confirmed that the OM may be applied to quantify parasite eggs in dehydrated sewage sludge containing polyelectrolytes.

Epidemiology of human toxocariasis in Poland - A review of cases 1978-2009.

Toxocariasis is a helminthozoonosis due to the infection of humans with larvae belonging to the Toxocara genus. Humans become infected as a result of accidental consumption of infected eggs containing third stage larvae (L3) nematodes from Toxocara canis or Toxocara cati species. Toxocariasis was recognized for the first time in the early 1950s, and the first cases of toxocariasis in Poland were described a few years later. Toxocariasis is clinically classified into several types: classic and incomplete visceral larva migrans (VLM) syndrome, ocular larva migrans (OLM) syndrome, neurological toxocariasis (NLM), covert toxocariasis and asymptomatic toxocariasis. In 1994-2005, 18,367 sera of people suspected of being infected with Toxocara were analysed, 1.8-76% had anti- Toxocara antibodies. In the period 1978-2009, 1,022 clinical cases of toxocariasis were recognized in Poland. In the opinion of the authors, in order to reduce the frequency of toxocariasis in human populations, some prophylaxis should undertaken, e.g. public education of zoonotic diseases, systematic control of animal, deworming of pets, cleaning pets' faeces by the owners.

Prevalence of antibodies against selected zoonotic agents in forestry workers from eastern and southern Poland.

INTRODUCTION AND OBJECTIVE: Due to the nature of their work, foresters are exposed to many infectious pathogens from the wildlife reservoir. The primary aim of this study was to assess their contact with these pathogens. MATERIAL AND METHODS: Sera and antibodies, mainly of IgG class against selected infectious factors, collected from a group of 216 employees of National Forests from eastern and southern Poland, were investigated. RESULTS: Antibodies to Anaplasma phagocytophilum were detected in 64 sera (29.6%), to Bartonella spp. in 133 sera (61.6%), to Babesia microti in 50 sera (23.1%), and to Coxiella burnetii in 14 sera (6.4%); 7 sera (3.2%) were positive to Francisella tularensis. In studies of diseases not carried by ticks positive results were found in 28 sera (12.9%) to Toxocara canis, 13 sera (6.0 %) to Trichinella spiralis, and in 7 sera (3.2%) to Echinococcus granulosus. Antibodies to Hanta viruses were detected with antigens from Puumala and Dobrava/Hantaan strains in both IgM and IgG class. In 9 foresters, positive results were found, including one positive result to 2 antigens, 5 to Dobrava/Hantaan antigen and 3 to Puumala antigen. CONCLUSION: Frequently higher percentages of positive results were found in people working utdoors, and in men when compared with women.

Assessment of viability of the nematode eggs (Ascaris, Toxocara, Trichuris) in sewage sludge with the use of LIVE/DEAD Bacterial Viability Kit.

Sewage sludges from wastewater treatment plants may contain live parasite eggs, which can be a source of humans and animals infection. According to the current rules, parasitological examination includes detection of the Ascaris spp., Trichuris spp. and Toxocara spp. eggs and estimation of their viability. The viability assessment based only on the incubation and observation of isolated egg is long and imprecise. The aim of this study was to develop sensitive and less labour-intensive methods for assessing viability of Ascaris spp., Toxocara spp. and Trichuris spp. eggs in sewage sludge. For this purpose, LIVE/DEAD Kit was used. Firstly, the possibility of distinguishing between live and dead eggs in water was assessed. Secondly, an appropriate amount of dyeing mixture needed to distinguish the live and dead eggs in the sewage sludge was determined using experimentally enriched samples and naturally contaminated samples of sludge. Eggs were isolated from the samples by own method which was a combination of flotation and sedimentation, preceded by a long mixing. After the last stage of the procedure, sediment containing the eggs of parasites was stained by LIVE/DEAD kit according to the manufacturer instructions, but with the use of different variants of dyes mixture concentration. The investigation showed that live and dead eggs of these three parasites could be differed by this method with the use of proper concentration of dyes. Live eggs were stained in green (Ascaris and Trichuris) and green-blue (Toxocara). However, all types of dead eggs were red coloured. The study demonstrated that after some modifications (resulted from the nature of the samples) the LIVE/DEAD kit is useful for assessing the viability of Toxocara, Ascaris and Trichuris eggs occurring in the sludge.

Contamination of vegetables, fruits and soil with geohelmints eggs on organic farms in Poland.

The objective of this study was to evaluate the contamination of vegetables, fruits and soil with zoonotic parasite eggs on organic and conventional farms in south-eastern Poland. To evaluate the contamination with eggs of zoonotic parasites, examinations were conducted on 8 conventional and 11 organic farms in south-eastern Poland from May-October in 2008 and 2009. The following fruit and vegetables were selected for the experiment: strawberry, leek, onion, carrot, zucchini, beetroot, parsley, potatoes, celery, rhubarb, lettuce, cabbage, broccoli, pumpkin, young beetroot leaves, cauliflower, French beans, turnip, fennel and sorrel. A total of 187 samples of vegetables, fruits and soil were examined by means of a modified flotation method according to Quinn et al. (1980). Contamination with Ascaris, Trichuris and Toxocara eggs was found, with a higher number of positive samples revealed on conventional (34.7%), compared to organic farms (18.9%). The level of contamination in soil samples from conventional farms was higher (88.5% positive samples), than of those from organic farms (32.8%). Of the 15 geohelmints eggs, positive samples were found in vegetables: 9 Toxocara eggs, 4 Ascaris eggs and 2 Trichuris eggs. No geohelmints eggs were observed in the strawberry samples. The consumption of vegetables and fruits contaminated with the eggs of parasites may be the cause of parasitoses in humans. Stricter sanitary standards on farms of all types may limit the incidence of parasitic zoonoses.

Contamination of soil with eggs of geohelminths in recreational areas in the Lublin region of Poland.

Recreational areas cover parks, bicycle paths, lawns, urban squares, sports complexes, holiday camp areas, playgrounds for children, beaches, and even spontaneously used green field open spaces. While using recreational areas, people take with them accompanying animals (dogs, cats). These animals constitute the main source and reservoir of many dangerous zoonoses, including parasitoses caused by roundworms of the genus Toxocara and Trichuris. The objective of the presented study was determination of the level of contamination of soil with parasites' developmental forms (eggs of Toxocara spp., Ascaris spp., Trichuris spp and Ancylostoma) in selected recreational areas in the Lublin Region of southeastern Poland. The material for the study was collected from beaches in recreational areas located around the Biale Lake near Wlodawa, the water reservoirs in Krasnobród and Janów Lubelski. The studies were carried out from May-October 2010 durng which time a total number of 215 samples of sand were collected. Eggs of Ascaris spp. and Trichuris spp. were detected based on the Polish Standard PN-Z-19000-4/2001. Beaches on the Biale Lake were the most contaminated. The eggs of intestinal parasites were isolated from 6 per 15 samples examined, which is 40%. In one sample, the presence of the eggs of 3 types of parasites were observed, while the eggs of Ancylostoma were not found in any of the samples. Attention should also be paid to the possibility of contamination of recreational areas with the eggs of intestinal parasites by wild animals. Recently in Poland, as well as in many other European countries, the phenomenon of synantropization of wild living animals has been observed, which most frequently concerned wild boar (Sus scrofa) and red fox (Vulpes vulpes).