Comparison of the tensile modulus of three 3D-printable materials used in dentistry.

BACKGROUND: Three-dimensional (3D) printing technology has brought much innovation to medicine and has been successfully adopted in many areas of dentistry. Although 3D printing techniques are being increasingly used, their advantages and disadvantages still need to be investigated, particularly with regard to the materials used in dentistry. Dental materials should be biocompatible and non-cytotoxic, and have sufficient mechanical integrity in the oral environment in which they are intended for use. OBJECTIVES: The present work aimed to identify and compare the mechanical properties of three 3D-printable resins. The materials included IBT Resin, BioMed Amber Resin and Dental LT Clear Resin. The Formlabs Form 2 printer was used. MATERIAL AND METHODS: A tensile strength test was performed on 10 specimens of each resin. Tensile modulus was measured on 2-millimeter-thick dumbbell-shaped specimens, 75 mm in length and 10 mm in width. The 10 specimens of each resin were mounted between the grips of a universal testing machine (Z10-X700). RESULTS: The results showed that BioMed Amber specimens cracked easily, yet no deformation was observed. The amount of force used to test the tensility of the specimens was the lowest for IBT Resin, while it was the highest for Dental LT Clear Resin. CONCLUSIONS: IBT Resin was the weakest material, whereas Dental Clear LT Resin was the strongest.

First report of Blastocystis sp. subtypes in natural water bodies in north-western Poland: a one-year monitoring.

Studies on the presence of Blastocystis subtypes in water samples are far less numerous compared with stool samples. The main aim of this study was to examine the occurrence of Blastocystis subtypes in 36 natural water bodies in north-western Poland in the period from winter 2009 to autumn 2010. Single PCR with the use of Blast 505-532/Blast 998-1017 set of primers was used to detect Blastocystis DNA in the obtained water samples. Sequencing of the obtained amplicons revealed the presence of ST1 and ST3 subtypes in five of the 36 (13.9%) examined water bodies within 1 year period. Further examinations with the use of new samples are needed in order to check if Blastocystis occurs in the examined water bodies at the present time, however, the risk of infection should be taken into consideration.

Colorectal cancer and Blastocystis sp. infection.

BACKGROUND: Blastocystis sp. is a common intestinal protozoan found worldwide. Based on gene analysis, 17 subtypes (STs, ST1-ST17) have been identified, 9 of which have been isolated from humans. Differences in clinical consequences may depend on differences among the STs. Here, we evaluated the prevalence of Blastocystis sp. in patients with colorectal cancer (CRC) compared to a control group and assessed the relationships between Blastocystis sp. infection and sex; age; and CRC grade, stage, and location. METHODS: The study included 107 CRC patients (41 women and 66 men, median age 65 years); 124 subjects without colorectal cancer or a history of oncological disease comprised the control group (55 women and 69 men, median age 63). Stool samples were collected from patients before oncological treatment and examined using light microscopy (iodine-stained smear). Additionally, PCR-based identification of Blastocystis sp. was performed in 95 stool samples from CRC patients and 76 stool samples from the control group. RESULTS: Light microscopy showed that the prevalence of Blastocystis sp. was significantly higher in CRC patients than in the control group (12.15% and 2.42%, respectively; p = 0.0041). Multivariate analysis showed that the odds of Blastocystis sp. infection were fivefold higher in the CRC group than in the control group. PCR-based molecular examinations demonstrated that the proportion of patients infected with Blastocystis sp. was significantly higher in the CRC group than in the control group (12.63% and 2.63%, respectively; p = 0.023). The predominant ST in the CRC group was ST3, detected in nine patients (75%), followed by ST1 (2 patients, 16.7%) and ST2 (1 patient, 8.3%). No association was found between Blastocystis sp. infection and age, sex, or CRC stage, grade, or location. CONCLUSIONS: The results showed that CRC was associated with an increased risk of opportunistic Blastocystis sp. infection, even before oncological treatment. To the best of our knowledge, this is the first report estimating the prevalence of Blastocystis sp. infection in CRC patients before oncological treatment in Europe.

The role of different species of wild ungulates and Ixodes ricinus ticks in the circulation of genetic variants of Anaplasma phagocytophilum in a forest biotope in north-western Poland.

The aim of this study was to reveal genetic variants of Anaplasma phagocytophilum strains occurring in different species of wild ungulates and in Ixodes ricinus ticks to check the role of the examined species in the circulation of the revealed variants in nature. The aim was also to determine if the detected variants of A. phagocytophilum are specific for particular game species as well as to examine their identity with other strains, including pathogenic ones. Sequences of the amplified groEL heat shock operon and msp2 gene fragments of A. phagocytophilum were obtained from samples collected between 2005 and 2007 from 14 roe deer (Capreolus capreolus), 13 red deer (Cervus elaphus), 1 fallow deer (Dama dama) and 4 wild boar (Sus scrofa) as well as 13 engorged and 11 questing I. ricinus ticks occurring in the area of Puszcza Wkrzanska Forest in north-western Poland. Analysis of the sequences showed the presence of five and four gene variants of groEL and msp2, respectively. The variants showed high identity with sequences derived from strains pathogenic to humans and/or domestic and companion animals. Cervids seem to play a more important role in the circulation of the detected variants in nature than wild boar. Some of the detected variants are not shared by roe and red deer. The results obtained on the basis of groEL and msp2 sequences are discrepant. Analysis of the groEL operon sequence provides more information on A. phagocytophilum strains than the msp2 gene sequence.

Acanthamoeba - pathogen and vector of highly pathogenic bacteria strains to healthy and immunocompromised individuals.

Acanthamoeba is a free-living protist pathogen, which is present in every place on Earth. 50 to 100 percent of the adult population has serum antibodies, specific for Acanthamoeba antigens. Acanthamoeba is an etiological agent of keratitis and encephalitis diagnosed in human. Acanthamoeba keratitis occurs in healthy persons and may lead to visual impairment and blindness, because corneal infection with this parasite fails to induce cell-mediated immune response due to the absence of resident antigen-presenting cells in the cornea. Systemic immunization with Acanthamoeba antigens induces Th1 cell-mediated immunity and serum IgG antibody, but do not prevent the development of keratitis. Immunization via mucosal surfaces stimulates IgA antibodies in tears and protects against the development of keratitis. Amoebae feed mainly on bacteria, fungi, and algae. By transferring intracellular bacteria, amoeba contributes to the spread of diseases dangerous to humans. Some microorganisms have evolved to become resistant to protist, since they are not internalized or able to survive, grow, and exit free-living protists after internalization. In many cases, the bacteria inside living amoebae survive longer, and multiply better, showing higher virulence. There is a hypothesis, which assumes that Acanthamoeba and symbiontic bacteria survive and multiply better in moist soil, rich in nitrogen compounds, particularly in the vicinity of the root systems of Alnus glutinosa, infected with nitrogen-fixing bacteria Frankia alni. Impact of soil environment created by nitrogen-fixing bacterium Frankia alni on specific relations between protists Acanthamoeba and highly pathogenic bacteria strains in Alnus glutinosa habitats in Poland continue to be established.

Molecular detecting of piroplasms in feeding and questing Ixodes ricinus ticks

The purpose of this study was to detect piroplasms, which are pathogens of veterinary and zoonotic importance in ticks, that were collected from ponies and field vegetation and to determine the role of Shetland ponies as potential reservoir hosts for piroplasms. A total of 1737 feeding and 371 questing Ixodes ricinus collected from horses or vegetation were tested for the presence of Babesia and Theileria DNA. Piroplasm 18S rRNA gene amplification was conducted, and the obtained amplicons were sequenced. Babesia DNA was detected in only three ticks (one tick collected from a pony and two collected from vegetation), and all of the obtained sequences had 100% similarity to B. divergens. Theileria DNA was not present in the examined ticks. Thus, the above results indicate that ponies are probably not essential hosts for the detected species of piroplasms. Piroplasm species typical for horses (Babesia caballi and Theileria equi) were not detected because I. ricinus is not their vector. The low infection rate of I. ricinus with B. divergens shows that the disease risk for the local horse population and people associated with pony horses is low, but it demonstrates their possible role as a source of human infection in northern Poland.

Molecular evidence for Toxoplasma gondii in feeding and questing Ixodes ricinus ticks.

The aim of the present study was to detect Toxoplasma gondii in ticks collected from ponies and field vegetation and to determine the role of Shetland ponies as a potential reservoir host for T. gondii. A total of 1737 feeding Ixodes ricinus collected from 49 horses and 371 questing ticks were tested by PCR and sequencing for the presence and genotyping of T. gondii. All ticks were examined in a previous study to detect and identify pathogenic bacterial species. The aim of this study was also to detect co-infection of ticks with these bacteria and T. gondii. Genotyping of the sequenced B1 gene revealed that detected T. gondii strains represented genotype I, which is pathogenic for humans. T. gondii genotype I was detected in 4.5% of all I. ricinus, including in 2.99% of feeding ticks and in 10.24% of questing ticks; this difference was statistically significant. Thus, the above results indicate that ponies probably are not an essential host for the detected sporozoan. Infections with more than one pathogenic species were rare and involved mostly T. gondii and Borrelia burgdorferi sensu lato. Our results confirmed the presence of T. gondii in I. ricinus and showed a new geographical habitat of T. gondii occurring in I. ricinus ticks in Poland.

Molecular evidence for bacterial pathogens in Ixodes ricinus ticks infesting Shetland ponies.

Ixodes ricinus has the potential to transmit zoonotic pathogens to humans and domestic animals. The feeding I. ricinus (n = 1737) collected from 49 Shetland ponies and questing ones from vegetation (n = 371) were tested for the presence and differentiation of the bacterial species. DNA of I. ricinus ticks was examined with PCR and sequencing analysis to identify species of Borrelia burgdorferi sensu lato (Bbsl), Anaplasma phagocytophilum and Rickettsia spp. Altogether, 24.3 % I. ricinus of the infested horses and 12.4 % ticks from vegetation carried at least one pathogen species. Horse-feeding ticks (19.2 %) were significantly more frequently infected with Borrelia spp. than questing ticks (4.8 %). Among Bbsl species, in I. ricinus infesting ponies, B. garinii, B. afzelii, B. burgdorferi sensu stricto, B. valaisiana and B. lusitanie and one species, B. miyamotoi related to relapsing fever group, were detected. The 73 flaB gene sequences of Borrelia obtained from feeding I. ricinus have been deposited in GenBank. Among Rickettsia species, two were identified: R. helvetica which was dominant and R. monacensis. Infections with more than one pathogenic species, involving mostly Bbsl and R. helvetica were detected in 6.3 % of infected ticks collected from horses. Shetland ponies may play an important role in the epidemiological cycle of Bbsl and probably could contribute to the natural cycle of A. phagocytophilum and R. helvetica as host for infected ticks. The awareness about these infectious agents in ticks from ponies might be an important criterion for the risk assessment of human diseases, especially as these animals are maintained for recreational purposes.

Molecular Characterization of Acanthamoeba spp. Occurring in Water Bodies and Patients in Poland and Redefinition of Polish T16 Genotype.

Acanthamoeba genus is divided into 20 genotypes (T1-T20) on the basis of the gene encoding 18S rRNA sequence. Using of at least 2 kbp gene fragments is strongly recommended to identify new genotypes and 5% difference is commonly used as a criterion of new genotypes, however, this value is questionable. In this paper, Polish Acanthamoeba strains described earlier on the basis of ~850 bp Ami fragment of 18S rRNA gene as T4, T11 and a new T16 genotype, have been analyzed using near-complete sequence of the gene. This analysis was needed because the Ami fragment does not reveal full variability within 18S rRNA gene. Phylogenetic analysis based on Ami fragment is biased by artifacts in the construction of the tree, so the fragment should not be used for identification of new putative Acanthamoeba genotypes. The analysis confirmed that the Polish sequences represent T4 and T11 genotypes and that the strains described earlier as T16 genotype are in fact a new subgroup of the T20 genotype and that this genotype should be divided into two subgroups: T20a (two strains described by [J. Eukaryot. Microbiol. 62 (2015) 69]) and T20b (11 Polish strains described in this study). The T20b subgroup was isolated from both clinical samples and water bodies used by people as bathing places and there is a risk of infection for humans during contact with water.

Molecular characterization of Cryptosporidium and Giardia occurring in natural water bodies in Poland.

Cryptosporidium and Giardia protozoa are zoonotic parasites that cause human gastroenteritis and can be transmitted to human through the fecal-oral route and water or food. Several species belong to these genera and their resistant forms occur in water, but only some of them are infectious to human. Health risk depends on the occurrence of infectious Cryptosporidium and Giardia species and genotypes in water, and only molecular techniques allow detecting them, as well as enable to identify the contamination source. In this work, genotyping and phylogenetic analysis have been performed on the basis of 18S rDNA and ß-giardin genes sequences of Cryptosporidium and Giardia, respectively, in order to provide the molecular characterization of these parasites detected earlier in five natural water bodies in Poland and to track possible sources of their (oo)cysts in water. Genotyping revealed a high similarity (over 99 up to 100 %) of analyzed sequences to cattle genotype of C. parvum isolated from cattle and human and to G. intestinalis assemblage B isolated from human. The sequences obtained by others originated from patients with clinical symptoms of cryptosporidiosis or giardiasis and/or with the infection confirmed by different methods. The contamination of three examined lakes is probably human-originated, while the sources of contamination of two remaining lakes are wild and domestic animals. Obtained phylogenetic trees support suggestions of other authors that the bovine genotype of C. parvum should be a separate species, as well as A and B assemblages of G. intestinalis.

Thermophilic potentially pathogenic amoebae isolated from natural water bodies in Poland and their molecular characterization.

The free-living amoebae (FLA) may live in the environment and also within other organisms as parasites and then they are called amphizoic. They are potentially pathogenic for humans and animals and are found in water that is a source of infection. The aim of this study was molecular detection and identification of these FLA in natural water bodies in North-Western Poland to evaluate the risk of the pathogenic amoebae infections. We examined surface water samples collected from 50 sites and first, the tolerance thermic test was performed in order to select thermophilic, potentially pathogenic strains. For molecular identification of FLA, regions of 18S rDNA, 16S rDNA and intergenic spacers were amplified. Acanthamoeba T4 and T16 genotypes of 18S rDNA gene and 18S rDNA of H. vermiformis were detected. We identified two variants of Acanthamoeba T4 genotype, two variants of Acanthamoeba T16 genotype and one variant of H. vermiformis. Identification of the T16 genotype and H. vermiformis in water was for the first time in Poland. Additionally, we made attempts to adapt the RLB method for detection and differentiation of FLA species and strains. PCR seems to be more sensitive than RLB hybridization, though.

PCR and real time PCR for the detection of Cryptosporidium parvum oocyst DNA.

Three DNA extraction kits were used, all without preliminary procedures, then DNA extraction was preceded with freeze/thaw cycles in three versions. A lack of desired effect resulted in the application of liquid nitrogen/water bath cycles before the use of the extractions in further experiments. The effectiveness of DNA extraction was measured by PCR signal and C(T) values of real time PCR. A comparison of the efficiency of various Cryptosporidium parvum undiluted oocyst treatments prior to DNA extraction with the use of three kits has shown that the best results were obtained after extraction of DNA with the QIAamp DNA Tissue Mini Kit (T kit), preceded by triple liquid nitrogen/water bath in 100 degrees C for 2 minutes and with overnight proteinase K digestion. After extraction with the T kit, the detection limit was 50 oocysts per 200 microl when effectiveness was evaluated with PCR and 10 oocysts in the case of real time PCR.

Molecular evidence of vector-borne pathogens coinfecting dogs from Poland.

Ticks of the genus Ixodes are vectors for many pathogens, including Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum and Rickettsia spp., and may also serve as vectors for Bartonella spp. However, the role of ticks in Bartonella transmission requires additional studies. The aim of this study was to investigate whether coinfection with two or more vector-borne pathogens can occur in the following three groups of dogs: I - dogs with suspected borreliosis (N = 92), II - dogs considered healthy (N = 100), and III - dogs with diagnosed babesiosis (N = 50). Polymerase chain reactions were performed to detect DNA of Anaplasma phagocytophilum, Rickettsia spp. and Bartonella spp. in the blood of dogs. In dogs of Group I, the DNA of both A. phagocytophilum and Bartonella sp. was detected (14% and 1%, respectively). In eight dogs, coinfection was indicated: A. phagocytophilum or Bartonella sp. with B. burgdorferi s.l. (the presence of antibodies against and/or DNA B. burgdorferi s.l.). In the case of five dogs positive for A. phagocytophilum DNA, no coinfection with B. burgdorferi s.l. was shown. In Group II, the DNA of A. phagocytophilum was detected in four dogs. In Group III, no pathogenic agents possibly transmitted by ticks were confirmed. No DNA of R. helvetica was detected in any of the groups studied.

[Biology, epidemiology and diagnostics of pathogenic waterborne protozoan parasites]. / Biologia, epidemiologia i diagnostyka chorobotwórczych pierwotniaków przenoszonych za posrednictwem wody.

Cryptosporidium, Giardia intestinalis, Cyclospora cayetanensis, Isosopra belli and micropsoridia are the most important and common pathogens found in humans and many other species of vertebrates. In humans, mainly in immunocompromised patients, children, pregnant women and elderly people, they are the most frequently identified protozoan parasites causing gastrointestinal disease worldwide. These pathogens have several transmission routes, including anthroponotic and zoonotic transmission. What is more, in many cases of epidemics caused by mentioned pathogens the major cause of infection was contaminated with these organisms water and food. In spite of many existing regulations of clearing and making use of drinking water supplies and recreational water, cosmopolitan protozoan parasites are still the danger of public health. These organisms are responsible for many waterborne outbreaks worldwide. Light microscopy and immunofluorescence assay have been used to identify these organisms in most laboratories. However, these traditional techniques have major limitations in the specific diagnosis, these methods are not sensitive enough to detect cysts or oocysts in environmental samples, so the new molecular tools must be applied. Recently, PCR-based techniques have been developed for detection and genetic characterization of the different species and population variants of protozoan parasites is central to the prevention, surveillance and control of gastrointestinal diseases. In this review were characterized biology, epidemiology and the progress in technology for detection and surveillance of the most important waterborne protozoan parasites.

[Bartonella spp. as a zoonotic pathogens transmitting by blood-feeding arthropods]. / Bartonella spp. jako patogeny odzwierzece przenoszone przez krwiopijne stawonogi.

Prior to 1993, Bartonella bacilliformis was the only member of the Bartonella genus. Now, the genus Bartonella currently contains over 30 species of Gram-negative bacteria that parasitize mammalian erythrocytes and endothelial cells. Bartonella spp. have been isolated from a variety of mammal species, most often from rodents, ruminants and carnivores, and these animals are implicated as reservoirs for the genus Bartonella. The persistent bacteriemia is more readily documented in the primary reservoir species and may occur less frequently or to a much lower lever in accidental hosts. In the natural host, clinical manifestations of the infection may be minimal or unrecognizable. Several insects have been implicated in Bartonella transmission, including flies and ticks. The reservoir host and vector varying depending on the Bartonella species involved, although, neither the reservoir, nor the vector has been identified definitively for many recently described Bartonella species. Humans are natural reservoir hosts for two species: Bartonella bacilliformis and Bartonella quintana, but many animal-associated Bartonella can also cause disease in humans. Members of the genus Bartonella are involved in a variety of human diseases, such as Carrion's disease, cat scratch disease, trench fever, bacillary angiomatosis, endocarditis, pericarditis and neuroretinitis. Most cases of bartonellosis are now diagnosed by tests based on PCR or through serological tests using specific antigens.

[Infestation of game animals from north-western Poland by common tick (Ixodes ricinus)]. / Infestacja wybranych gatunków zwierzat lownych z pólnoc- no-zachodniej Polski przez kleszcza pospolitego (Ixodes ricinus) (Acari, Ixodida, Ixodidae).

BACKGROUND: Ixodes ricinus tick species is the most common tick in Poland and is the primary vector of many pathogens. The aim of this study was to determine the infestation of three species of game animals: roe deer (Capreolus capreolus), red deer (Cervus elaphus) and wild boar (Sus scrofa) from forest area of north-western Poland. MATERIAL AND METHODS: Examined ticks have been collected from roe deer (Capreolus capreolus), red deer (Cervus elaphus) and wild boar (Sus scrofa). RESULTS: Single specimen of roe deer harboured from 1 to 9 ticks (mean value 3,6), red deer - from 1 to 8 ticks (mean value 4,4) and wild boar - from 1 to 3 ticks (mean value 2,3). Prevalence of ticks at roe deer was 44,2%, red deer - 40 % and wild boar - 6%. Statistical analysis revealed that difference in tick prevalence between roe deer and red deer is not significant in contrast to difference between roe deer and wild boar and between red deer and wild boar, so, most likely, prevalence of tickborne pathogens is larger in examined ruminants than in wild boar. Examined species (especially ruminants) play important role in maintaining of tickborne pathogens in a local habitat, not only as a potential reservoir, but also as hosts of I. ricinus ticks.

Wild game as a reservoir of Anaplasma phagocytophilum in north-western Poland.

BACKGROUND: The aim of the study was to determine the role of game animals as reservoirs of Anaplasma phagocytophilum, a bacteria species transmitted by Ixodes ricinus ticks, from north-western Poland (Zachodniopomorskie vovoidship). The area under question is endemic for A. phagocytophilum. MATERIAL AND METHODS: Blood and spleen samples were taken from 72 roe deer between April and December 2003. Animals culled during winter did not harbour ticks, on the other hand 155 individuals of Ixodes ricinus were collected from 35 of 43 animals taken during spring. We tested all samples for A. phagocytophilum by PCR amplification of the msp2 gene. An individual was considered infected if pathogens were detected in at least one isolate (blood or a tissue sample). RESULTS: DNA from A. phagocytophilum was not found in isolates from ticks collected from the animals. The general level of infection for the roe deer was 31.94% (23/72). DNA of A. phagocytophilum was most commonly detected in blood samples; only in three cases was anaplasma DNA detected in spleen and not in blood. Ruminants seem to be the most competent reservoir for A. phagocytophilum in north-western Poland.

Prematurity and protracted mechanical ventilation as risk factors for Pneumocystis jiroveci infection in HIV-negative neonates in an intensive care unit.

This work was undertaken to elucidate some aspects of the epidemiology of Pneumocystis pneumonia (PP). We studied 42 mechanically ventilated, human immunodeficiency virus (HIV)-negative, severely ill neonates treated at an intensive care unit. The study group included 40 premature neonates and two mature neonates with lethal congenital defects. Progressive respiratory dysfunction in PP necessitated mechanical ventilation. Infection was usually noticeable on the 22nd day of life or after 12 days of ventilation. The usual manifestations included apnea, pallor, copious frothy sputum, seizures, and feeding difficulties. The diagnosis was established by detecting Pneumocystis jiroveci cysts in bronchial lavage fluid specimens (88.1% sensitivity). PP was managed with cotrimoxazole and pentamidine combination therapy administered over 14 days. No clinical improvement was noted in four neonates and three of them died during therapy. Prematurity and protracted mechanical ventilation are two risk factors for P. jiroveci infection in severely ill neonates in an intensive care unit.

PCR detection of granulocytic Anaplasma and Babesia in Ixodes ricinus ticks and birds in west-central Poland.

The aim of the study was to establish the role of forest birds as reservoirs of Anaplasma phagocytophilum and Babesia spp. in Wielkopolski National Park. A total of 108 birds from 9 species were collected between May-September 2002. Blood samples were taken from 84 specimens and 442 individuals of the common tick, Ixodes ricinus, were collected from the birds. The 73 additional ticks were collected from vegetation. PCR amplification of a fragment of the epank 1 gene and 18S rRNA gene was used for detection of A. phagocytophilum and Babesia spp. DNA, respectively. Pathogen DNA was not detected in any of the blood samples or ticks collected from birds. On the other hand, 3 ticks collected from vegetation (4.1% of all examined specimens) were positive for A. phagocytophilum DNA. In spite of the high level of infestation of birds by I. ricinus, it is clear that they do not constitute a competent reservoir of A. phagocytophilum and Babesia in WNP. Additionally, I. ricinus is not a significant vector in this area.

[Roe deer (Capreolus capreolus) and red deer (Cervus elaphus) as a reservoir of protozoans from Babesia and Theileria genus in north-western Poland]. / Sarny (Capreolus capreolus) i jelenie (Cervus elaphus) jako rezerwuar pierwotniaków z rodzaju Babesia i Theileria w pólnocno-zachodniej Polsce.

The species of genus Babesia and Thelieria are obligate intracellular pathogens that multiply in both vertebrate and invertebrate hosts. Some species of Babesia cause bovine babesiosis infecting erythrocytes of the cattle and wild ruminants, and undergo a complex developmental cycle in ticks which serve as biological vectors. Majority of Theileria spp. cause bovine theileriosis infecting lymphocytes as well as erythrocytes of the cattle and wild ruminants, and similar to Babesia undergo a complex developmental cycle in ticks. In this study, hunter killed roe deer (Capreolus capreolus) and red deer (Cervus elaphus) from north-western Poland were tested for Babesia and Theileria infection in two seasons (spring and autumn, 2004). Infection with babesias and theilerias was detected by PCR assay based on the fragment of nuclear small subunit rRNA gene (nss-ribosomal DNA). Four types of products different in size were obtained and then sequenced. Sequence analysis of nucleotides showed that two kinds of products (385 and 475 bp) were unspecific, the third was characteristic for Theileria sp. (430bp) and the last one for Babesia divergens (407bp). We found that 24.4% of the animals examined were infected with Babesia divergens and 11% with Theileria sp. Percentage of infected animals with B. divergens was almost equal in the spring and autumn (24.6 and 24% respectively). Infection with Theileria was lower in the spring than in the autumn (10.5 and 12% respectively).