Introduction: The aim of the study was to test the utility of Flinders Technology Associates filter paper (FTA® Cards) for molecular detection and storage of avian influenza virus (AIV). Material and Methods: There were two strains of AIV used in the study: low pathogenicity H7N1 and high pathogenicity H5N1 subtypes. Detection of viral material was conducted using molecular RT-PCR and rRT- PCR method. Results: The infectivity of LPAIV/H7N1 and HPAIV/H5N1 was completely inactivated within 1 h and 24 h after adsorption to FTA® Cards at room temperature, respectively. Viruses stored on FTA® Cards had detection limit approximately 1 log₁₀ lower than live viruses. Viral RNA of both strains were detectable on the cards by rRT-PCR for a minimum of 150 d, irrespectively of storage temperatures (room temperature, -20ºC). RNA was also detected in all samples obtained from SPF chickens experimentally infected with HPAI/H5N1 on 3ʳᵈ and 4ᵗʰ day post-infection (p.i.). Conclusion: FTA® Cards enable safe and effective alternative transport of samples for molecular diagnosis of AIV.

Introduction: The aim of the study was to test the utility of Flinders Technology Associates filter paper (FTA® Cards) for molecular detection and storage of avian influenza virus (AIV). Material and Methods: There were two strains of AIV used in the study: low pathogenicity H7N1 and high pathogenicity H5N1 subtypes. Detection of viral material was conducted using molecular RT-PCR and rRT- PCR method. Results: The infectivity of LPAIV/H7N1 and HPAIV/H5N1 was completely inactivated within 1 h and 24 h after adsorption to FTA® Cards at room temperature, respectively. Viruses stored on FTA® Cards had detection limit approximately 1 log10 lower than live viruses. Viral RNA of both strains were detectable on the cards by rRT-PCR for a minimum of 150 d, irrespectively of storage temperatures (room temperature, -20ºC). RNA was also detected in all samples obtained from SPF chickens experimentally infected with HPAI/H5N1 on 3rd and 4th day post-infection (p.i.).

Introduction: The aim of the experiment was to establish the enterotoxigenic Escherichia coli K88 (ETEC K88)-induced BALB/c mouse duodenum inflammation model. Material and Methods: Mice were administered different concentrations of E. coli K88 (1.0 × 10⁷-10⁹ CFU/mL) for 3 d by means of an esophageal catheter. Results: The results showed that the treated group expressed several significant clinical symptoms, such as reduced dietary demands and weight loss, an increased presence of IL-1α, TNF-α, and MPO in the peripheral blood, and some pathological changes in the duodenum. On the 6ᵗʰ-8ᵗʰ days, the body weight of the mice was the lowest. On the 8ᵗʰ day, there were significant differences in IL-1α, TNF-α, and MPO levels compared to the control group (P < 0.05), the gap between the duodenum mucous layer and the muscular layer had widened, the number of goblet cells was increased, and the inflammatory infiltrate and inflammation changes in the lamina propria and the mucous layer were the most obvious. Conclusion: The duodenum inflammation was the most severe on day 8; thus, the model was successfully established. In addition, varying concentrations of ETEC K88 did not significantly influence the duodenum inflammation (P > 0.05).

Introduction: The aim of the experiment was to establish the enterotoxigenic Escherichia coli K88 (ETEC K88)-induced BALB/c mouse duodenum inflammation model. Material and Methods: Mice were administered different concentrations of E. coli K88 (1.0 × 107-109 CFU/mL) for 3 d by means of an esophageal catheter. Results: The results showed that the treated group expressed several significant clinical symptoms, such as reduced dietary demands and weight loss, an increased presence of IL-1α, TNF-α, and MPO in the peripheral blood, and some pathological changes in the duodenum. On the 6th-8th days, the body weight of the mice was the lowest. On the 8th day, there were significant differences in IL-1α, TNF-α, and MPO levels compared to the control group (P < 0.05), the gap between the duodenum mucous layer and the muscular layer had widened, the number of goblet cells was increased, and the inflammatory infiltrate and inflammation changes in the lamina propria and the mucous layer were the most obvious. Conclusion: The duodenum inflammation was the most severe on day 8; thus, the model was successfully established. In addition, varying concentrations of ETEC K88 did not significantly influence the duodenum inflammation (P > 0.05).

Introduction: Due to the high heterogeneity of canine lymphoma, the aim of the present study was to test in vitro the chemosensitivity of canine high-grade primary lymphoma cells to various cytostatic drugs commonly used to treat dogs: 4-HO-cyclophosphamide, doxorubicin, dexamethasone, prednisolone, vincristine, etoposide, chlorambucil, lomustine, and cytosine arabinoside. Material and Methods: To determine the cell viability and drug ability to induce apoptosis two different tests were used: an MTT assay and annexin V/propidium iodide staining. Results: Both in vitro tests were found to be useful tools. Significant differences in the sensitivity, depending on the drug type, between B-, T- and mixed/null-type lymphoma cells were found for the majority of the tested drugs. B-type cells were the most sensitive in vitro, whereas T-type cells seemed to be the most resistant. Doxorubicin, chlorambucil, etoposide, and vincristine most strongly reduced the cell viability and induced apoptosis. Conclusion: In vitro assays, such as the MTT test and especially the annexin V/PI assay, may be useful tools for predicting a response to the treatment of high-grade lymphoma in dogs or improving the treatment outcomes in individual animals.

Introduction: Due to the high heterogeneity of canine lymphoma, the aim of the present study was to test in vitro the chemosensitivity of canine high-grade primary lymphoma cells to various cytostatic drugs commonly used to treat dogs: 4-HO-cyclophosphamide, doxorubicin, dexamethasone, prednisolone, vincristine, etoposide, chlorambucil, lomustine, and cytosine arabinoside. Material and Methods: To determine the cell viability and drug ability to induce apoptosis two different tests were used: an MTT assay and annexin V/propidium iodide staining. Results: Both in vitro tests were found to be useful tools. Significant differences in the sensitivity, depending on the drug type, between B-, T- and mixed/null-type lymphoma cells were found for the majority of the tested drugs. B-type cells were the most sensitive in vitro, whereas T-type cells seemed to be the most resistant. Doxorubicin, chlorambucil, etoposide, and vincristine most strongly reduced the cell viability and induced apoptosis. Conclusion: In vitro assays, such as the MTT test and especially the annexin V/PI assay, may be useful tools for predicting a response to the treatment of high-grade lymphoma in dogs or improving the treatment outcomes in individual animals.

Introduction: The differentially expressed proteins between healthy cows and those with footrot were identified to explore changes in protein profiles associated with the disease. Material and Methods: Out of 36 cows selected for the experiment, 18 footrot-affected cows were included in the treatment group (group T) and 18 unaffected cows were included in the control group (group C). Plasma samples from groups T and C were subjected to two-dimensional electrophoresis analysis and differentially expressed proteins were identified by matrix-assisted laser desorption/ionisation tandem time-of-flight mass spectrometry. Bioinformatics, including gene ontology analysis and pathway analysis, was used for analysing all proteins. Results: Out of 63 spots identified by 2DE, 33 were selected for mass spectrum analysis, which identified 11 differentially expressed proteins in 26 spots. Footrot led to changes in profiles in plasma proteins that were classified to the pathway of inflammatory response, complement, and blood coagulation, among others. Conclusion: This study provides evidence of the defence mechanisms of cows with footrot to explore strategies for treatment.

Introduction: The differentially expressed proteins between healthy cows and those with footrot were identified to explore changes in protein profiles associated with the disease. Material and Methods: Out of 36 cows selected for the experiment, 18 footrot-affected cows were included in the treatment group (group T) and 18 unaffected cows were included in the control group (group C). Plasma samples from groups T and C were subjected to two-dimensional electrophoresis analysis and differentially expressed proteins were identified by matrix-assisted laser desorption/ionisation tandem time-of-flight mass spectrometry. Bioinformatics, including gene ontology analysis and pathway analysis, was used for analysing all proteins. Results: Out of 63 spots identified by 2DE, 33 were selected for mass spectrum analysis, which identified 11 differentially expressed proteins in 26 spots. Footrot led to changes in profiles in plasma proteins that were classified to the pathway of inflammatory response, complement, and blood coagulation, among others. Conclusion: This study provides evidence of the defence mechanisms of cows with footrot to explore strategies for treatment.

There are numerous biomarkers of central and peripheral nervous system damage described in human and veterinary medicine. Many of these are already used as tools in the diagnosis of human neurological disorders, and many are investigated in regard to their use in small and large animal veterinary medicine. The following review presents the current knowledge about the application of cell-type (glial fibrillary acidic protein, neurofilament subunit NF-H, myelin basic protein) and central nervous system specific proteins (S100B, neuron specific enolase, tau protein, alpha II spectrin, ubiquitin carboxy-terminal hydrolase L1, creatine kinase BB) present in the cerebrospinal fluid and/or serum of animals in the diagnosis of central or peripheral nervous system damage in veterinary medicine.

There are numerous biomarkers of central and peripheral nervous system damage described in human and veterinary medicine. Many of these are already used as tools in the diagnosis of human neurological disorders, and many are investigated in regard to their use in small and large animal veterinary medicine. The following review presents the current knowledge about the application of cell-type (glial fibrillary acidic protein, neurofilament subunit NF-H, myelin basic protein) and central nervous system specific proteins (S100B, neuron specific enolase, tau protein, alpha II spectrin, ubiquitin carboxy-terminal hydrolase L1, creatine kinase BB) present in the cerebrospinal fluid and/or serum of animals in the diagnosis of central or peripheral nervous system damage in veterinary medicine.

Introduction: The paper describes identification of Trichinella species isolated from wild boars (Sus scrofa) in the most popular hunting region of the West Pomeranian Province of Poland.Material and Methods: The Trichinella larvae were identified by digestion method. For species identification of the larvae, multiplex PCR was used according to the European Reference Laboratory for Parasites Multiplex PCR protocol. The results were confirmed by molecular amplification of 5S rDNA gene and sequence analysis.Results: Prevalence of 0.54% Trichinella–positive wild boars in the West Pomeranian Province was recorded. Examination of the larvae showed the occurrence of T. spiralis in 79 %, T. britovi in 16.5 %, mixed infection with T. spiralis/T. britovi in 3.5%, and T. pseudospiralis in 1.0% of the boars.Conclusion: This is the first record of wild boar infected with non-encapsulated larvae of T. pseudospiralis in Poland. The species is very difficult to determine, especially using trichinoscopic method. The discovery of the larvae in the animals which may be intended for human consumption confirms that digestion technique should be the only method used for the inspection of meat, especially that from wild boars..

Introduction: The paper describes identification of Trichinella species isolated from wild boars (Sus scrofa) in the most popular hunting region of the West Pomeranian Province of Poland.

Introduction: The article presents a rapid and simple analytical procedure for determination of four tetracyclines (oxytetracycline, tetracycline, chlortetracycline, and doxycycline) in animal medicated feedingstuffs. Material and Methods: Two-gramme samples were extracted by a Na₂EDTA-McIlvaine buffer (pH 4)/methanol mixtures (40/60, v/v). The determination was achieved by liquid chromatography using a Zorbax Eclipse XDB C18 analytical column with mass spectrometer detection (LC-MS). Results: Recoveries of the antibiotics from spiked feed samples ranged from 78.2% to 113.5%. The LOD and LOQ for tetracyclines in feeds ranged from 2.8 to 4.2 and from 4.3 to 5.7 mg/kg, respectively. Conclusion: The method was successfully validated and proved to be efficient, precise, and useful for quantification of tetracyclines in medicated feedingstuffs.

Introduction: The article presents a rapid and simple analytical procedure for determination of four tetracyclines (oxytetracycline, tetracycline, chlortetracycline, and doxycycline) in animal medicated feedingstuffs. Material and Methods: Two-gramme samples were extracted by a Na2EDTA-McIlvaine buffer (pH 4)/methanol mixtures (40/60, v/v). The determination was achieved by liquid chromatography using a Zorbax Eclipse XDB C18 analytical column with mass spectrometer detection (LC-MS). Results: Recoveries of the antibiotics from spiked feed samples ranged from 78.2% to 113.5%. The LOD and LOQ for tetracyclines in feeds ranged from 2.8 to 4.2 and from 4.3 to 5.7 mg/kg, respectively. Conclusion: The method was successfully validated and proved to be efficient, precise, and useful for quantification of tetracyclines in medicated feedingstuffs.

Introduction: This article presents the analysis of the correlation between the category and health status of calves and the results of their rearing and levels of selected blood parameters.Material and Methods: The study included 105 Polish Holstein-Friesian and beef (Limousine, Charolaise and Hereford) crossbred calves. Young bulls were purchased at the age of two to four weeks. The animals underwent quarantine, were dehorned, and 46 young bulls were castrated. The germ horns were removed by burning out. Castration was carried out with a bloodless method using a rubber band. The calves were kept in groups and fed a milk replacer administered via teats from automated milk-feeding stations. After the period of milk feeding, the calves were fed grass silage ad libitum and a concentrate at 2.5 kg/animal/day. The calves were weighed every two weeks. Blood for analyses was sampled at 43 d of age.Results: After the rearing period finished at the age of six months, young bulls and steers had similar body weights (176.17 and 176.55 kg) and approximate average daily weight gains from birth (0.756 and 0.767 g/day). The healthy calves at six months of age weighed 180.47 kg, whereas the animals which at least once suffered from some diseases during rearing were lighter by approx. 30 kg (P ≤ 0.01). A statistically significant (P ≤ 0.01) difference was found for the count of red blood cells and white blood cells. In comparison with healthy individuals, the diseased animals had less RBC (8.33 and 9.42 10¹²/L respectively) and more WBC (27.03 and 12.26 10⁹/L respectively).Conclusion: Castration of young bulls did not have any impact on the results of rearing and health status of the calves. The magnitude of the analysed parameters depended on the health status of the calves. Thus RBC and WBC parameters may be used to predict the health status of calves during rearing.

Introduction: This article presents the analysis of the correlation between the category and health status of calves and the results of their rearing and levels of selected blood parameters.

Introduction: The main goal of the study was to investigate the effect of KATP channel modulators on development of oxidative stress in the heart of rats showing different resistance to hypoxia.Material and Methods: The study has been performed on rats showing high- (HR) or low-resistance (LR) to hypoxia under modulators of ATP-sensitive potassium (KATP) channel opener pinacidil (0.06 mg/kg) and blocker glibenclamide (1 mg/kg) upon cobalt (Co) treatment (30 mg of cobalt chloride/kg b.w., 3 h). Changes in the oxidative stress parameters of the heart tissue, such as lipid peroxidation (LPO), level of oxidatively modified protein (OMP), and antioxidant defence system (superoxide dismutase - SOD, catalase -CAT, glutathione peroxidase - GPx, glutathione reductase - GR) as well as total antioxidant activity (TAA) were analysed.Results: Co treatment caused a significant decrease in SOD and CAT activity in the heart of LR rats and GPx activity in HR rats. It also led to a decrease in OMP level in the heart of rats with HR in comparison with controls.Conclusion: The obtained results suggest that individual resistance to hypoxia plays a crucial role in Co actions and provides evidence that the effects of KATP channel opener pinacidil in the heart are mediated through different pathways of the antioxidative system, depending on the individual resistance to hypoxia. Pinacidil exerts a protective effect on the heart tissue by preventing the LPO decrease and significantly reducing OMP levels, as well as increasing TTA in rats with LR.

Introduction: The main goal of the study was to investigate the effect of KATP channel modulators on development of oxidative stress in the heart of rats showing different resistance to hypoxia.

Introduction: The study examined the concentration of total mercury and correlation coefficients between fish size or FCF (condition factor) and the content of Hg in muscle tissue of six freshwater fish: bream (Abramis brama L.), roach (Rutilus rutilus L.), whitefish (Coregonus lavaretus L.), vendace (Coregonus albula L.), perch (Perca fluviatilis L.), and pike (Esox lucius L.). Material and Methods: The fish were caught from the Lake Pluszne located in the Olsztyn Lake District (Poland). Mercury was analysed by atomic absorption spectrometry using Milestone DMA-80 (with dual-cell). Results: The content of the element in the muscles of the examined fish was as follows: pike (0.197 mg/kg) ≈ perch (0.173 mg/kg) > vendace (0.114 mg/kg) ≈ roach (0.095 mg/kg) and roach ≈ whitefish (0.065 mg/kg), and whitefish ≈ bream (0.042 mg/kg) (p ≤ 0.05). In all cases, the content of mercury correlated positively with the body weight and total length of the fish. Only the correlation coefficients between mercury concentration and weight or length of bream were slightly higher (0.979 and 0.977 respectively, p ≤ 0.001). The length and weight relationship of the fish was also determined. Conclusion: The results showed that the levels of mercury were lower than the maximum acceptable limit established by the Commission Regulation (EC) No 629/2008 of 2 July 2008. Thus, they are safe from consumer health point of view.

Introduction: The study examined the concentration of total mercury and correlation coefficients between fish size or FCF (condition factor) and the content of Hg in muscle tissue of six freshwater fish: bream (Abramis brama L.), roach (Rutilus rutilus L.), whitefish (Coregonus lavaretus L.), vendace (Coregonus albula L.), perch (Perca fluviatilis L.), and pike (Esox lucius L.). Material and Methods: The fish were caught from the Lake Pluszne located in the Olsztyn Lake District (Poland). Mercury was analysed by atomic absorption spectrometry using Milestone DMA-80 (with dual-cell). Results: The content of the element in the muscles of the examined fish was as follows: pike (0.197 mg/kg) ≈ perch (0.173 mg/kg) > vendace (0.114 mg/kg) ≈ roach (0.095 mg/kg) and roach ≈ whitefish (0.065 mg/kg), and whitefish ≈ bream (0.042 mg/kg) (p ≤ 0.05). In all cases, the content of mercury correlated positively with the body weight and total length of the fish. Only the correlation coefficients between mercury concentration and weight or length of bream were slightly higher (0.979 and 0.977 respectively, p ≤ 0.001). The length and weight relationship of the fish was also determined. Conclusion: The results showed that the levels of mercury were lower than the maximum acceptable limit established by the Commission Regulation (EC) No 629/2008 of 2 July 2008. Thus, they are safe from consumer health point of view.