Comparison of serum protein profiles of Borrelia burgdorferi-positive Bernese Mountain Dogs and dogs of other breeds using MALDI-TOF technique.

The aim of the study was to compare the serum protein profile of Bernese Mountain Dogs (BMDs) reacting positive for Bb in snap testing with the serum protein profile of dogs of other breeds (healthy and with clinical borreliosis) using the MALDI time-of-flight (MALDI-TOF) technique. The observations included five groups of dogs. BMDs reacting positively to Bb in snap serological testing and showing symptoms of borreliosis (group 1), BMDs for which no borreliosis symptoms were determined but with seropositivity for Bb determined with snap serological tests (group 2), clinically healthy BMDs with no antibodies for Bb found in the serum (group 3), five dogs of different breeds, reacting positively in serological testing, in which borreliosis symptoms were observed (group 4), clinically healthy dogs of different breeds with negative reaction in tests towards Bb (group 5). A proteomic analysis demonstrated the presence of five identical protein fractions among all five groups. An additional two protein fractions of approximately 7.630 and 15.260 kDa were found in all the serum samples obtained from the dogs positive for borrelia in a snap test, both in those exhibiting symptoms of borreliosis, and seropositive BMDs not presenting symptoms of the disease. These two additional protein fractions may be used to differentiate between seropositive and seronegative B. burgdorferi dogs and may be considered a seropositivity marker, however, it cannot be used to differentiate between animals with the clinical form of the disease and those that are only seropositive.

Lyme disease in Bernese Mountain Dogs. Is it a real problem?

Borreliosis is the most frequently diagnosed tick-borne disease caused by spirochete bacteria belonging to the genus Borreliae - Borrelia burgdorferi sensu stricto (s.s.), Borrelia afzelii and Borrelia garinii. Clinical manifestations in dogs include fever, lameness, polyarthritis and glomerulonephritis. Diagnosis is mainly serological and is based on an immunoenzymatic test followed by a Western blot confirmatory test. Early treatment with antibiotics such as doxycycline or amoxicillin, for four weeks, usually reduces the risk of chronic disease. Tick control, including tick repellents, is highly reliable in preventing transmission. Vaccines are available to reduce transmission and the clinical manifestations of infection in dogs. Bernese Mountain Dogs are a breed that often test positive for antibodies against B. burgdorferi without showing any clinical symptoms of the disease. Quantitative determination of the immunoglobulin level for spirochetes has indicated that Bernese Mountain Dogs may have an increased susceptibility to Borrelia spp. infections of a hereditary nature.

Clinical course of granulocytic anaplasmosis in hunting dogs.

The aim of this study was to analyze cases of granulocytic anaplosmosis diagnosed in 53 hunting dogs in Poland. Medical records of dogs naturally infected with Anaplasma phagocytophilum were retrospectively evaluated with regard to clinical signs and laboratory abnormalities at the time of presentation, therapy and course of disease. The most common clinical signs in A. phagocytophilum-positive dogs included in the study were lethargy (100%), inappetence (94%) and fever (92.5%). Thrombocytopenia was the most common laboratory abnormality (100%), followed by a drop in haematocrit level (79.3%) and increased AST activity (75.5%). Of the 53 infected dogs, 51 (96%) recovered and two dogs (with neurological symptoms) died. Analysis of these cases indicates that A. phagocytophilum infection must be considered in differential diagnosis in dogs living in Poland, especially in hunting dogs with thrombocyto- penia and Ixodes ricinus tick invasions.

Babesia gibsoni infection in dogs in Poland.

Canine babesiosis is a tickborne, protozoal, haemoparasitic disease. Babesia organisms are frequently classified as either large (B. canis) or small (B. gibsoni). The aim of this study was an attempt to detect B. gibsoni DNA in blood samples taken from dogs suspected of suffering from tick-borne diseases. 216 samples were tested using PCR, of which, in 99 of them B. canisDNA was detected, whereas in 3 of them B. gibsoni was detected. Positive PCR results for B. gibsoni were confirmed using a Qube MDx real-time analyzer. The results indicate that infec-tions with this B. gibsoni should be taken into account and included in the differential diagnosis of vector-borne diseases in dogs in Poland, and that the accurate identification of the species of parasite causing the infection is crucial for developing the correct treatment regimen and prognosis.

Application of Matrix-assisted Laser Desorption Ionization Time-of-flight Mass Spectrometry for Identification of Coagulase-negative Staphylococci Isolated from Milk of Cows with Subclinical Mastitis.

The aim of this study was to use matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) for the identification of coagulase-negative staphylococci (CNS) isolated from the milk of cows with subclinical mastitis. The study material consisted of 33 isolates of CNS, identified by the results of API Staph tests, obtained from the milk of cows with subclinical mastitis. Based on the spectra analyses, MALDI-TOF MS tests of 33 bacterial samples allowed identification of the microorganisms in 27 cases (81.8%). The most frequent cause of subclinical mastitis was found to be Staphylococcus sciuri (39%), while S. vitulinus was detected in 15% of the milk samples. The results obtained indicate that MALDI-TOF MS can be used for the identification of CNS isolated from bovine mastitis as a method supplementary to biochemical tests.

Genome-wide co-localization of active EGFR and downstream ERK pathway kinases mirrors mitogen-inducible RNA polymerase 2 genomic occupancy.

Genome-wide mechanisms that coordinate expression of subsets of functionally related genes are largely unknown. Recent studies show that receptor tyrosine kinases and components of signal transduction cascades including the extracellular signal-regulated protein kinase (ERK), once thought to act predominantly in the vicinity of plasma membrane and in the cytoplasm, can be recruited to chromatin encompassing transcribed genes. Genome-wide distribution of these transducers and their relationship to transcribing RNA polymerase II (Pol2) could provide new insights about co-regulation of functionally related gene subsets. Chromatin immunoprecipitations (ChIP) followed by deep sequencing, ChIP-Seq, revealed that genome-wide binding of epidermal growth factor receptor, EGFR and ERK pathway components at EGF-responsive genes was highly correlated with characteristic mitogen-induced Pol2-profile. Endosomes play a role in intracellular trafficking of proteins including their nuclear import. Immunofluorescence revealed that EGF-activated EGFR, MEK1/2 and ERK1/2 co-localize on endosomes. Perturbation of endosome internalization process, through the depletion of AP2M1 protein, resulted in decreased number of the EGFR containing endosomes and inhibition of Pol2, EGFR/ERK recruitment to EGR1 gene. Thus, mitogen-induced co-recruitment of EGFR/ERK components to subsets of genes, a kinase module possibly pre-assembled on endosome to synchronize their nuclear import, could coordinate genome-wide transcriptional events to ensure effective cell proliferation.

Liquid Storage of Hematopoietic Stem Cells Versus Proliferative Potential Colony-Forming Unit Granulocyte-Monocytes: Validation of Cell Processing.

BACKGROUND: The material for transplantation must be of the highest quality. As far as we know, short-term storage is one of the crucial points of stem cell banking. According to the quality assurance system in a stem cell bank, each step of cell processing must be validated. The aim of this study was to assess the influence of short-term storage conditions into a clonogenic assay. METHODS: Material was collected from mobilized peripheral blood by means of leukapheresis from 15 patients. Samples were stored at 4°C and 20°C; samples were evaluated on the day of leukapheresis and after 24 hours and after 48 hours of storage. The number of colony-forming unit granulocyte-monocyte (CFU-GM) precursors was analyzed with the use of in vitro culture. The material was evaluated before freezing and after thawing. RESULTS: The average number of CFU-GM precursors in the material stored at 4°C before freezing on the day of collection was 84/10(5) nuclear cells (nc) and after 24 hours and 48 hours of storage was, respectively, 62/10(5) nc (P = .011719) and 36/10(5) nc (P = .02088). The average of the CFU-GM precursors in material stored at 20°C after 24 hours and 48 hours of storage amounted to 33/10(5) nc (P = .004439) and 2/10(5) nc (P = .00346), respectively. CONCLUSIONS: In our study, the number of colonies of CFU-GM after 24 hours and 48 hours of storage, both at 4°C and 20°C, was significantly reduced compared with the number of colonies on the day of collection. Significantly greater numbers of CFU-GM precursors were observed in the material stored before freezing at 4°C in comparison with the material stored at 20°C.

Banking of Hematopoietic Stem Cells: Influence of Storage Time on Their Quality Parameters.

BACKGROUND: Banking of hematopoietic stem cells (HSCs) is a rapidly growing part of the transplant field. The essence of the banking process is to maintain the optimal quality parameters throughout the storage period, allowing successful transplantation. METHODS: Our laboratory research was carried out on 126 HSC samples that were collected by means of leukapheresis from patients with lymphoproliferative diseases. The samples were frozen in a controlled rate and stored up to 76 months in containers in vapor phase of liquid nitrogen. The evaluation was performed after thawing the probes. Viability of nuclear cells was assessed after incubation in Trypan blue, CD34+ phenotype cells were determined by means of cytometry with the use of 7-aminoactinomycin D (7-AAD), and an analysis of the proliferative potential of granulocyte-monocyte precursors was performed. For comparative statistical analysis, the material was divided into 3 groups according to storage time: A: <1 month (n = 45); B: 1-12 months (n = 50); C: >12 months (n = 31). RESULTS: In the examined groups, similar median values were observed of nuclear cell viability (A, 86%; B, 87%; and C, 83%) and CD34+ cells (95%, 94.5%, and 95.8%, respectively). A gradual, nonsignificant, reduction in the median of granulocyte-monocyte precursors was found: 68 × 10(4)/kg of body weight (kg bw), 48.5 × 10(4)/kg bw, and 47 × 10(4)/kg bw, respectively. Statistical analysis with the use of the Kruskal-Wallis test showed a P value of >.05 for all variables. CONCLUSIONS: There were no significant differences in the viability of nuclear cells, CD34+ cells, and proliferative potential granulocyte-monocyte precursors between groups. Storage for up to 76 months does not change the essential quality parameters, and HSCs could be qualified for distribution.

Helmintofauna of turbot Scophthalmus maximus (Linnaeus, 1758) from the southern Baltic Sea including new data.

Turbot Scophthalmus maximus (Linnaeus, 1758) is a fish belonging to the Pleuronectiformes order. It is commonly observed in waters of the northern Atlantic, and also in the Baltic Sea. As an economically significant species, it is fished on an industrial scale, and also farmed in some European countries. Seventy-two turbots from the Gulf of Gdansk (26th ICES zone) were examined for parasite presence in the years 2010-2012. The study revealed the presence of the tapeworm Bothriocephalus scorpii (Müller, 1776) and acanthocephalan Corynosoma semerme (Forssell, 1904). The overall (both parasites) prevalence of turbot infection was 100% with a mean intensity of 18.7. C. semerme is a parasite which has not been noted so far in turbot from the southern Baltic. The presence of C. semerme in turbot was emphasized in the context of possible infection of terrestrial mammals, including humans.

CD55, CD59, factor H and factor H-like 1 gene expression analysis in tumors of the ovary and corpus uteri origin.

The expression level of complement regulators in ovarian and corpus uteri tumors was not fully established so far. In current manuscript we performed gene expression analysis by the real-time PCR approach to investigate both membrane bound - CD55 and CD59 and fluid phase - factor H and factor H-like 1 complement regulators. We found increased CD55 expression in corpus uteri tumors when compared to control tissues, whereas in ovarian cancer CD55 expression was lower than in control sections. Additionally we found CD59 expression to be more prominent in ovarian cancer than in corpus uteri tumor samples. We observed also the strong positive correlation between the level of expression of the whole group of regulators, which was particularly significant between the expression of factor H and factor H- like 1. In conclusion we present novel results which implicates different role of particular complement inhibitors in the regulation of the complement system in two cancer types examined. Strong positive correlation between examined proteins implicates similar pattern of the regulation which should be taken into consideration with regards to the possible immunotherapy applied as adjuvant therapeutic approach in these two indications. The inhibition of complement regulation may serve as a strategy to potentiate the efficacy of such treatment.

Analysis of the effect of cryoprotectant medium composition to viability of autologous hematopoietic cells collected by leukapheresis.

OBJECTIVES: Cryopreservation of hematopoietic stem cells intended for autologous transplantation is a crucial element of the banking process. Although cryopreservation techniques are well known, improvement is needed. This study was designed to optimize cryopreservation to improve the quantitative and qualitative parameters of hematopoietic stem cells in the material intended for transplantation. We used available opportunities to provide the best quantitative and qualitative parameters of hematopoietic stem cell transplants processed in a closed system. MATERIAL AND METHODS: Two hundred forty-eight products of hematopoietic stem cells collected by leukapheresis from patients with lymphoproliferative disorders create the basis of this report. The material was frozen in a controlled-rate freezer and stored in containers in the vapor phase of LN2 (-160°C). The composition of a cryoprotectant medium was modified. For freezing, 192 probes were used with a cryoprotective medium containing 20% dimethyl sulfoxide (DMSO) and enriched RPMI 1640. For 56 samples, we used 20% DMSO in autologous plasma harvested during leukapheresis. Products of hematopoietic stem cells and cryoprotectant medium were combined in a 1:1 ratio. The final number of nuclear cells did not exceed 2 × 10(8)/mL. Analysis was performed after thawing the probes. Viability of nuclear cells has been assessed using the microscopic technique after incubation in Trypan blue and the CD34+ cells by flow cytometry using the 7-aminoactynomycin D. A statistical analysis has been conducted using the Statistica program (StatSoft, Cracow, Poland). RESULTS AND CONCLUSIONS: The results show that the application of autologous plasma is linked with higher viability of nuclear cells and CD34+ cells. Moreover, statistical analysis of the nuclear cells and CD34+ cells viability differs significantly between groups frozen using RPMI 1640 and autologous plasma (P < .05). To assess the viability of CD34+, cells frozen using RPMI 1640 results showed a large span of at 16.4% to 99.1% living cells.

Microbial contamination of peripheral blood and bone marrow hematopoietic cell products and environmental contamination in a stem cell bank: a single-center report.

Hematopoietic stem cells (HSC) derived from peripheral blood (PB) and bone marrow (BM) are frequently used for autologous and allogenic transplantations. Establishing quality control at appropriate steps of the stem cell preparation process is crucial for a successful transplantation. Microbial contamination of haematopoietic stem cells is rare but could cause a potentially mortal complication of a stem cells transplantation. We investigated the microbiological contamination of PB (291 donations) and BM (39 donations) products. Microbial cultures of 330 donations between January 2012 and June 2013 were retrospectively analyzed after the collection and preparation steps. The microbiological analysis was performed with an automated system. Hematopoietic stem cells were processed in a closed system. Additionally, in this report the environment of the working areas of stem cell preparation was monitored. We analyzed microbial contamination of the air in a class I laminar air flow clean bench at the time of preparation and in the laboratory once per month. We reported 9 (2.73%) contaminated HSC products. The most frequent bacteria isolated from PB and BM products were Bacillus species. Coagulase-negative staphylococci and Micrococcus species were the most frequent micro-organisms detected in the air microbial control. Microbial control results are necessary for the safety of hematopoietic stem cell products transplantation. Microbial control of hematopoietic stem cell products enables an early contamination detection and allows for knowledgeable decision making concerning either discarding the contaminated product or introducing an efficient antibiotic therapy. Each step of cell processing may cause a bacterial contamination. A minimum of manipulation steps is crucial for increasing the microbial purity of the transplant material. Also, the air contamination control is essential to ensure the highest quality standards of HSC products preparation.

Associations of physical activity and inactivity with body tissue composition among healthy Polish women and women after mastectomy.

The aim of the study is to determine the relationship between physical activity and body composition among healthy women and women who have had mastectomy. This is in order to establish whether physical activity of women after breast cancer treatment improves composition and distribution of body mass components to a degree which will allow to achieve the body composition of healthy women. Research material consists of anthropometric measurements (body height, weight) of women and bioelectric impedance analysis (BIA) of body composition (using Akern - BIA 101 composition analyzer). Intensity of activity was assessed using the Physical Activity International Questionnaire. The sample consisted of 76 healthy women (active 44.74%, inactive 55.26%) and 70 females after mastectomy (54.29% and 45.71%, respectively). Mean chronological age of women after mastectomy was 53.40 years, SD=7.55, and of the healthy ones 52.38 years SD=11.01). A significant difference in body composition was noted among active and inactive women after mastectomy; namely the active females had lower weight (by approximately 12 kg), body mass index (BMI), level of fat mass (by approximately 8%) and (by approximately 5%) total body water. The active healthy women had 6% less fat mass, almost 4% more body water and 6% more fat free mass. Programmed physical activity undertaken by women after mastectomy is recommended and produces good results in the form of reduction of excessive body fat tissue. Through physical activity these women are able to achieve the same level of fat mass as healthy women.

Detection of Borrelia burgdorferi sensu lato DNA in the blood of wild bison from Bialowieza primeval forest in eastern Poland.

The aim of the present study was to investigate the occurrence of Borrelia burgdorferi sensu lato DNA in a group of 120 wild bison (Bison bonasus) from the Bialowieza Primeval Forest in eastern Poland. The PCR technique revealed the presence of 16S RNA of Borrelia burgdorferi sensu lato in the blood of 16 out of 120 examined animals. DNA amplification by means of primers SC1 and SC2 gave a product with a size of 300-bp. The sequences of the PCR products obtained showed 100% homology with each other and 100% homology with B. burgdorferi s.1. 16S RNA gene DQ111061. Results of this study suggest that wild bison are important in maintaining agents of Lyme borreliosis, and that studies of reservoir competence of this species are indicated.

Nuclear estrogen receptors co-activation mechanisms.

Estrogens play very important role in opening the transcription event, which is a final step of activation of the first order mediators as receptors or channels in the cell wall by information coming from the outside of the cell. For the long time the exact step by step mechanism of cellular transfer of information to the cell nuclei was not known. Currently many new informations are available. Very important seems the step of phosphorylation and therefore desensitization of the target proteins. All peptide kinases, especially serine and threonine, like protein kinases A and C, RAS and MAP kinases, cycline kinases are potential or confirmed biological targets. Except them elements of the transcription complexes like p160.SRC-1, histon acetyltransferase and histon deacetylase, CBP/p300, TRAP/DRIP, NSD1, PPARγ/PGC-1, NCOR1, SMRT, REA were also found useful. Finally estrogens are able to activate other receptors, namely aryl hydrocarbon receptors (AhR) and estrogen receptor related proteins (ERR). It is well known that many types of cancer are related to the direct or indirect excessive activation of nuclear estrogen receptors, therefore their inhibition could be crucial in many estrogen-related cancers. Understanding the interactions in such complexes would help in developing new and better multi-target cures and finding new ligands with better pharmacological and pharmacokinetic properties.

Molecular analysis of a fragment of gene E1B 19K of canine adenovirus 2 (CAV-2) isolated from dogs with symptoms of cough.

The aim of this study was to perform molecular analysis of canine adenovirus 2 (CAV-2) E1B 19K gene fragment isolated from 20 dogs of various breeds (12 males and 8 females aged 1-9 years), with clinical symptoms of upper respiratory tract infections, from the Lubelszczyzna region. Nasal swabs were taken from dogs. DNA of CAV-2 was detected using the PCR method in 16 swabs. All PCR products were sequenced, and the obtained sequences were compared with each other and with the sequence of the E1B 19K gene of the CAV-2 strain from an online database of NCBI GenBank: AC 000003. Based on analysis of the obtained sequences, three polymorphic variants of CAV-2 (No. 1-3) with homology of 78 - 100% were distinguished. The nucleotide and amino acid sequences of the most frequently represented polymorphic variant, No. 1, differed from the sequences of polymorphic variant No. 2 with one substitution. The nucleotide and amino acid sequence of the E1B 19K gene of CAV-2 AC 000003 differed from the analogous sequences of representatives of variant No. 1 with 44 nucleotide and 19 amino acid substitutions. The small number of nucleotide differences in the E1B 19K CAV-2 gene among the examined own isolates, compared with AC 000003, suggest that the infections in dogs were caused by a relatively genetically stable virus which occurs in eastern

The identification of Anaplasma spp. isolated from fallow deer (Dama dama) on a free-range farm in eastern Poland.

The aim of the present study was to investigate the occurrence of Anaplasma spp. in group of 50 fallow deer (Dama dama) from free-range farm in eastern Poland and determine what species of Anaplasma could infect these animals based on PCR gene sequencing. The PCR technique revealed the presence of 16S RNA Anaplasma spp. genetic material in the blood of 7 out of 50 examined animals. The sequences of the PCR products obtained showed a 100% homology with each other and 100% homology with GU 183908 sequence of A. phagocytophilum, isolated in our earlier study from a horse with clinical form of anaplasmosis. Here, we report the first molecular evidence of Anaplasma spp. among naturally infected fallow deer in eastern Poland.

Multiple trichoepitheliomas--a novel mutation in the CYLD gene.

BACKGROUND: Trichoepitheliomas are benign neoplasms with follicular differentiation. They may present as a solitary lesion or as multiple lesions. Multiple trichoepitheliomas are inherited in an autosomal dominant pattern within families, with both variable penetrance and expressivity. Recent investigations support that mutations in CYLD, the gene affected in familial cylindromatosis as well as in Brooke-Spiegler syndrome, are also responsible for multiple trichoepitheliomas. OBJECTIVE: The authors report the case of a 9-year-old African girl with multiple facial trichoepitheliomas in whom a mutation in the CYLD gene was hypothesised. MATERIALS AND METHODS: After genomic DNA extraction from the peripheral blood, a molecular analysis of the CYLD gene was performed by PCR, DHPLC and automated sequencing. RESULTS: A novel heterozygous mutation in exon 18 of the CYLD gene (c.2449delT) was identified, with a deletion of one nucleotide resulting in a premature translational termination codon at amino acid position 831 on the affected allele (p.Cys817Valfs X15). CONCLUSIONS: The predominating tumours define the classification of these three entities. Nevertheless, studies suggest that they can simply represent phenotypic variations of the same disease spectrum, sharing common genetic mutations.

Factors affecting neutrophil recovery after autologous bone marrow-derived stem cell transplantation in patients with acute myeloid leukemia.

Bone marrow is currently regarded as the most appropriate source of stem cells for patients with acute myeloid leukemia (AML) undergoing autologous transplants. A total of 55 adult patients with AML in first complete remission receiving autologous bone marrow-derived stem cell (BMSC) transplantation (BMSCT) were analyzed to determine factors affecting the rate of neutrophil recovery. All patients were treated with standard induction and three to four courses of consolidation chemotherapy and, after collection of BMSC, conditioned with BuCy2. The median time to neutrophil reconstitution was 30 (10-62) days and was delayed in 24 patients. Neutrophil recovery was faster in patients who had received granulocyte-macrophage progenitors (CFU-GM) dose >23.5 x 10(4)/kg, CD34(+) cells >3.2 x 10(6)/kg, and mononuclear cells (MNCs) >3 x 10(8)/kg. The speed of neutrophil recovery correlated with the number of transplanted CFU-GM progenitors (P = .0077) and MNCs (P = .0015). CFU-GM progenitors dose was the only factor close to significance in univariate analysis of neutrophil engraftment. Probability for neutrophil recovery was higher in patients transplanted with a higher dose of MNCs. These data suggested that the content of CFU-GM progenitors and MNCs within the bone marrow graft was the most important factor for the quality of neutrophil recovery after autologous BMSCT in AML patients.

The clinical course of babesiosis in 76 dogs infected with protozoan parasites Babesia canis canis.

The aim of the study was to trace the clinical course of babesiosis in 76 dogs infected with Babesia canis protozoa and to assess the usefulness of PCR method in the routine diagnosis of the disease. The investigations were conducted in three successive seasons of the biological activity of ticks on dogs displaying possible clinical signs of babesiosis, the latter assigned individual numbers from 001 to 076. All the animals underwent routine clinical examinations and blood was collected for haematological, biochemical, parasitological and molecular tests for babesiosis. The most frequent clinical signs observed in the course of the disease were changes in urine colour and xanthosis or paleness of mucous membranes, whereas in the haematological and biochemical examinations, the most frequent laboratory findings were thrombocytopenia, leucopoenia, anaemia and an increase in levels of urea and bilirubin. In all blood smears stained with the May-Grunwald and Giemsa methods, from the 76 dogs, the presence of Babesia canis protozoa was observed in erythrocytes, and their DNA was detected in 69 blood samples by means of PCR technique. The course of the disease and the results of molecular examinations suggested the haemolytic form of babesiosis. The previous genetic analysis of isolates of Babesia canis canis from the eastern areas of Poland helped to distinguish two specific groups, A and B, within the species (Adaszek and Winiarczyk 2008a). The present study revealed a certain interrelation between the intensification of thrombocytopenia and the fact that protozoa belong to either group A or B. The mean number of thrombocytes in dogs infected with protozoa from group A was 61.11 thousand/mm3, whereas the mean number of thrombocytes in the blood of dogs infected with protozoa from group B was 27.47 thousand/mm3. A strong correlation was also observed between the low level of thrombocytes and the increase in the internal body temperature (p = 0.02), accelerated pulse rate (p = 0.01) and discoloration of urine (p = 0.04). As a result of the treatment of dogs with imidocarb, recovery was observed in 73 out of the 76 dogs brought to the clinic.