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Purpose The European wild boar (Sus scrofa) is a popular game animal species. Its meat, however, can represent a reservoir of dangerous foodborne diseases and can play an important role in the transmission of many pathogens, including Toxoplasma gondii, in humans and animals worldwide. The aim of the present study was to determine the presence of antibodies to T. gondii in the serum of hunted wild boars in Poland. Methods Using the commercial direct agglutination test, 398 serum samples collected during the hunting season 2009/2010 were tested for the presence of T. gondii antibodies, and the titre of 40 was considered indicative of T. gondii infection in analysed samples. Results It was found that nationwide, 37.7% were seropositive to T. gondii, although seroprevalence varied from 11.6 to 50% depending on the Voivodeship. Significant differences were observed between the Great Poland and Lubusz Voivodeships and between Great Poland and Warmian-Masurian. Conclusion Serological test indicated widespread exposure to T. gondii by wild boar; therefore, consumption of raw or undercooked game meat of infected animals can carry a significant risk of T. gondii infection.

Rodents play an important role as reservoir hosts of zoonotic diseases. As a component of our long-term programme of monitoring parasitic infections in bank vole populations in three ecologically similar sites in NE Poland, we screened blood samples for signs of a serological response to the presence of Trichinella spp. The overall seroprevalence of Trichinella spp. was 1.52%, but prevalence was largely concentrated in one of our three study sites and confined to the oldest individuals in the study. Seroprevalence of Trichinella spp. did not differ between the sexes. Although a local prevalence of 1.52% may seem low, when this is extrapolated to the national population of bank voles in peak years, perhaps numbering hundreds of millions of animals, the number of infected bank voles on a country wide scale is likely to be huge. Our results suggest that bank voles may be reservoirs of Trichinella spp. However, on the basis of our results we consider their importance as epidemiologically significant hosts for Trichinella spp. to be moderate and their role in this context to require further investigation., Graphical abstract Image 1, Highlights • Seroprevalence of Trichinella spp. was relatively stable between the years of study. • There was significant variation of Trichinella spp. seroprevalence between study sites maintained over nine years study. • Trichinella spp. seroprevalence was highest in the oldest bank voles and was essentially identical in both sexes.

Ashworthius sidemi, a blood-sucking abomasal nematode, has been identified in various wild ruminants, including deer (Cervus elaphus), roe deer (Capreolus capreolus), fallow deer (Dama dama) and moose (Alces alces). Although it has been observed throughout Poland, most sightings have been in the eastern part of the country. However, more recently, A. sidemi has been confirmed in the Ruszów Forest District (Lower Silesian Wilderness). It is now possible to test the faeces of cattle for the DNA of the third-stage infectious larvae (L3) of A. sidemi. The present paper describes such a molecular study of 120 faecal samples collected from cattle grazed in the Ruszów Forest District and Biebrza Marshland, where A. sidemi had previously been detected in wildlife. In this study, no A. sidemi DNA was identified in any of the examined samples.

The studies were carried on raccoons from Poland, Germany and the Czech Republic. Tissue samples from raccoon hearts, lungs and brains were used for molecular examination while meat juice was collected for immunological tests. Antibodies against T. gondii were detected in six out of 44 raccoons (13.6%), while T. gondii DNA was found in 18 (40.9%). Antibodies against N. caninum were found in seven raccoons (15.9%) but no parasite DNA was observed in any sample. DNA of T. gondii was observed in raccoons of both sexes (in 42.3% of females and 38.9% of males) from all three countries. The proportion of raccoons that tested positive for DNA of T. gondii was higher in the Czech Republic (47.1%) than in Germany (33.3%), however the difference was non-significant (p = 0.7032). It seems that the raccoons appear to have been exposed to both T. gondii and N. caninum, but only T. gondii infection was confirmed. The role of raccoons as reservoir, and as possibly contributing to spread of these parasites merits further studies.

Toxoplasma gondii, a coccidian parasite known to infect almost all warm-blooded animals, is the cause of one of the most common zoonotic parasitic diseases. The aim of the study is to determine whether the 529 bp fragment or the TGR1E gene is more useful target for PCR identification ofT. gondii, for common use. The brains of 221 carnivores and omnivores collected between 2013 and 2015 from north-eastern Poland were examined for the presence of this parasite. The DNA was extracted and then amplified using specific primers. Positive results were obtained in 24% of brain samples using the TGR1E target and 19% using the 529 bp sequence. The results demonstrate that both TGR1E and 529 bp repeat element are suitable for detectingT. gondiiDNA in wildlife animals, and the combination of two methods is necessary to obtain reliable results.

Toxoplasma gondii and Neospora caninum are coccidian parasites with a global distribution that cause reproductive failure and production losses in livestock. The seroprevalence of both parasite species in ruminants and Cervidae has been investigated worldwide and found to vary greatly. Studies carried out on mixed flocks with 3 ruminant species (sheep, goats, and fallow deer) living under the same conditions are excellent models for identifying any differences in the rate of infection with the 2 parasites between the animal species. Additionally, the species used in the present study differ in their feeding categories: grazers, browsers, and intermediate feeders. The aim of the study is to identify any variation in the prevalence of the 2 parasites in mixed flocks and to identify any possible relationships with food choice. The seroprevalence against T. gondii and N. caninum in 167 captive fallow deer, 64 sheep, and 39 goats were detected using commercially available ELISA. The seroprevalence for T. gondii achieved 10% in fallow deer, 21% in goats, and 47% in sheep. The seroprevalence for N. caninum achieved 13% in sheep and fallow deer and 21% in goats. Overall, 53% of the sheep, 33% of the goats, and 22% of the fallow deer were seropositive for both infections. Coinfection of T. gondii and N. caninum was detected in 6% of sheep, 8% of goats, and 2% of fallow deer. Statistical analyses of the seroprevalence levels observed between 2 parasites for each animal species revealed that only the results obtained for sheep were significant (P < 0.01). Additionally, the differences in the seroprevalence levels for T. gondii between sheep and goats and between sheep and fallow deer were statistically significant (P < 0.01). The results of the N. caninum seroprevalence levels observed among animal species were not significant. Although the variations in susceptibility to T. gondii and N. caninum infections demonstrated by the examined animals may affect the differences in seropositivity, these appear to be related to the feeding habits of the animal species. Therefore, the risk of infection by agents found close to the ground, such as coccidian oocysts, varies. Sheep as grazers are at a greater risk of infection by T. gondii than goats and fallow deer.

The study was performed on a male European bison (Bison bonasus bonasus L.) foetus spontaneously aborted at the fourth or fifth month of pregnancy in the Bialowieza Forest. Serum samples from the foetus and mother revealed the presence of antibodies against T. gondii (S/P% = 88% and 75%, respectively). Mobile extracellular tachyzoites were first observed in a Vero cell culture, 110 days following inoculation of brain homogenate. PCR amplification with TGR1E1 and TGR1E2 primers confirmed the presence of T. gondii DNA, which was classified as Type I by PCR-RFLP genotyping. The sequences of 18S ribosomal RNA (18S rRNA) and 5.8S ribosomal RNA (5.8S rRNA) genes; internal transcribed spacer 1 (ITS1) and internal transcribed spacer 2 (ITS2), obtained from T. gondii isolate, have been deposited in GenBank (accession number KX459518.1). This is the first in vitro isolation and molecular identification of T. gondii from an aborted European bison foetus. The origin of this protozoan isolate indicates that the species is a significant threat to the European bison conservation program implemented in the Bialowieza Forest.

The raccoon (Procyon lotor) is a species native to North America, but which is now spreading throughout Europe. Raccoons have been found to host various Trichinella species. The aim of the present study was to evaluate the effectiveness of using immunological testing of meat juice for determining the occurrence of Trichinella in raccoons. The studies were carried out on 139 animals from three European countries: the Czech Republic, Germany and Poland. Seven meat juice samples were found to be positive for antibodies to Trichinella by ELISA, and another seven were unclear. The ELISA results were confirmed by immunoblot: anti-Trichinella antibodies were identified in 9.35 % of the examined animals. Slight agreement (κ = 0.13) was found between the digestion method and the combined ELISA and immunoblot approach. From the results of our study, we concluded that meat juice may be used as a simple and convenient sample for detection of anti-Trichinella in racoons.

The raccoon (Procyon lotor) is a North American carnivore introduced to Europe in the 20th Century. Raccoons are believed to be the potential hosts of many parasites, or to be involved in their transmission to other animals. Nematodes of the genus Trichinella can infect many carnivorous and omnivorous animals worldwide. The aim of the present study was to determine the occurrence of Trichinella spp. infection in raccoons in Central Europe. Muscle samples were collected from various regions of Poland, the Czech Republic and Germany during the years 2012-2016. The larvae of Trichinella spp. were detected in 11 raccoons, and these were identified as T. spiralis and T. pseudospiralis by multiplex PCR (89.9% and 9.1%, respectively). No mixed infection was observed. This is the first report describing the occurrence of T. spiralis and T. pseudospiralis in P. lotor in Central Europe. Our findings also show that the raccoon population acts as a reservoir of Trichinella pseudospiralis.

Trichinellosis is one of the most widespread parasitic zoonoses. Trichinella Owen, 1835 nematodes are found in pigs, horses, and humans in the domestic cycle, and in many carnivores and omnivores in the sylvatic cycle, such as wild boars, red foxes, raccoon dogs, and wolves. Carnivores are known to be involved in the circulation of Trichinella nematodes and they act as a reservoir in the sylvatic environment. The aim of this study was to determine the occurrence of Trichinella spp. infection in red foxes in Poland. Samples were collected from 2010 to 2015 in different regions of the country and then tested for Trichinella nematodes using HCl-pepsin digestion. Trichinella larvae were found in 10.02% of examined samples (145/1447). The larvae were identified as T. spiralis (11.03%), T. britovi (71.72%), and T. pseudospiralis (0.69%). No mixed infection was observed. The prevalence of infection varied between years and different voivodeships of the country. Our findings confirm that red foxes are involved in the maintenance of Trichinella spp. in the sylvatic cycle in Poland.

The aim of the study was to compare the usefulness of two antibody-based methods, the direct agglutination test (DAT) and enzyme linked immuosorbent assay (ELISA), with that of the polymerase chain reaction (PCR) for detecting anti-Toxoplasma gondii in samples derived from naturally-infected wild animals. Antibodies against T. gondii were detected in meat juice samples collected from 129 free- living carnivores and omnivores. T. gondii seroprevalence was confirmed in 73,6% of examined samples when DAT and ELISA were used separately, but in only 88,4% samples when both immunological tests were used in parallel. PCR results confirmed the presence of DNA of the parasite in 24 of all the 129 samples. Sixteen samples were classified as positive when all three tests were used. A moderate degree of agreement was found between DAT and ELISA (κ=0.55). However, no agreement was found between the molecular and serological tests: κ=-1.75 for DAT versus PCR; κ=-1.67 ELISA versus PCR. By using both serological tests, antibodies against T. gondii were found in 77.5% of red foxes, 12.5% of badgers, 40% of martens and 8.3% of raccoon dogs. Antibodies against the parasite were detected also in one mink, but not in the sample derived from a polecat. T.gondii DNA was found in the brain tissue of 20 red foxes, three badgers and one raccoon dog. Our studies confirm that ELISA and DAT are suitable and reliable techniques for T. gondii antibody detection in meat juice from wild animals when serum samples are unavailable. Positive results obtained by immunological tests do not always reflect that the host was infected by T. gondii. They indicate only a contact with parasite. PCR should be used to confirm te presence of DNA from T. gondii.

A simple polymerase chain reaction (PCR) test was used to identify Ashworthius sidemi , a blood-sucking gastrointestinal nematode that commonly infects bison, red and roe deer, and moose in Poland. The present study uses this technique to confirm the possibility of transmission of A. sidemi infection from wildlife to domestic animals, such as cattle and sheep, grazing on the same natural pastures. A 406 bp fragment of genomic A. sidemi DNA was actually detected in DNA isolated from larval cultures derived from feces from cattle. A. sidemi DNA has been detected in cattle which represent a new host for this parasite. This is the first evidence of A. sidemi in cattle. The results reveal that a PCR test based on DNA from L3 larvae can be used for in vivo detection of A. sidemi invasions in breeding animals. In conclusion, the transfer of A. sidemi infection from wildlife to the farm animals sharing the same pastures appears possible.

Trichinellosis is an epidemiological problem with a global distribution. In Poland a substantial increase of the wild boar population has been observed since 2010, together with an increased incidence of trichinellosis after ingestion of raw or undercooked wild boar products containing Trichinella spp. larvae. However, the actual number of human cases remains particularly difficult to determine. The aim of the present study was to determine the current prevalence and spread of these parasites within wild boars. The diaphragm pillars and tongue from 833 wild boars were collected from 2010 to 2014, as well as one wild boar meat sausage known to be a source of infection. The samples were tested for Trichinella spp. using pepsin digestion. Recovered larvae were identified at species level by multiplex polymerase chain reaction (multiplex PCR). The overall prevalence in all examined samples was found to be 2.0% (17/833). Recovered larvae were identified as T. spiralis and T. britovi (9/18 and 5/18, respectively). T. spiralis larvae were isolated from the sausage. Mixed infection was confirmed only once. Three isolates were not identified. The results of our study confirm that the wild boar plays a key role in the maintenance of Trichinella nematodes through the sylvatic cycle.

Several penicillin biosensors obtained by covalent binding of penicillinase directly to the surface of pH detectors are presented. Two types of pH electrodes were used: membranes with tridodecylamine, as hydrogen ionophore, incorporated in carboxylated polyvinyl chloride, and membranes made of aminated polyvinyl chloride without any ionophore. Short response time and high stability characterize all described biosensors. The effects of several parameters on the shape of the calibration curves of the biosensors are discussed.

Neospora caninum Dubey, Carpenter, Speer, Topper et Uggla, 1988 is a protozoan parasite originally reported as a major cause of bovine abortions worldwide. It is documented that the parasite is widely spread among non-carnivorous cervids. The purpose of this study was to investigate the seroprevalence of N. caninum in moose (Alces alces Linnaeus). Blood samples collected in 2010 and 2012 in the northeastern Poland were tested for antibodies to N. caninum by agglutination test (NAT), a commercial competitive screening enzyme-linked immunosorbent assay (cELISA) and enzyme-linked immunoassay (ELISA). Sera that gave a positive result were further investigated by western blot (WB) analysis to verify the presence of antibodies. Antibodies to N. caninum were detected in one of seven moose. The antibody titer was confirmed by NAT (1 : 1 280), cELISA (I = 91%) and ELISA (OD = 0.736). The main immunodominant antigens detected by WB were 120, 70, 55, 35 and 16 kDa proteins. This is the first evidence of N. caninum seropositivity in moose living in a natural environment in Europe.