Introduction: The genomes of nine H5 subtypes of low pathogenic avian influenza virus (LPAIV) strains identified in wild birds in Poland between 2010 and 2015 were sequenced, and their phylogenetic relationship was determined.

Introduction: The genomes of nine H5 subtypes of low pathogenic avian influenza virus (LPAIV) strains identified in wild birds in Poland between 2010 and 2015 were sequenced, and their phylogenetic relationship was determined. Material and Methods: AIV genome segments were amplified by RT-PCR and the PCR products were sequenced using Sanger method. Phylogenetic trees were generated in MEGA6 software and digital genotyping approach was used to visualise the relationship between analysed strains and other AIVs. Results: High genetic diversity was found in the analysed strains as multiple subgroups were identified in phylogenetic trees. In the HA tree, Polish strains clustered in two distinct subclades. High diversity was found for PB2, PB1, PA and NP, since 5-8 sublineages could be distinguished. Each strain had a different gene constellation, although relationship of as much as six out of eight gene segments was observed between two isolates. A relationship with poultry isolates was found for at least one segment of each Polish strain. Conclusion: The genome configuration of tested strains indicates extensive reassortment, although the preference for specific gene constellation could be noticed. A significant relationship with isolates of poultry origin underlines the need for constant monitoring of the AIV gene pool circulating in the natural reservoir.

Introduction: The prevention and control of Actinobacillus pleuropneumoniae in commercial production settings is based on serological monitoring. Enzyme-linked immunosorbent assays (ELISAs) have been developed to detect specific antibodies against a variety of A. pleuropneumoniae antigens, including long-chain lipopolysaccharides (LPS) and the ApxIV toxin, a repeats-in-toxin (RTX) exotoxin unique to A. pleuropneumoniae and produced by all serovars. The objective of this study was to describe ApxIV antibody responses in serum and oral fluid of pigs.

Introduction: The prevention and control of Actinobacillus pleuropneumoniae in commercial production settings is based on serological monitoring. Enzyme-linked immunosorbent assays (ELISAs) have been developed to detect specific antibodies against a variety of A. pleuropneumoniae antigens, including long-chain lipopolysaccharides (LPS) and the ApxIV toxin, a repeats-in-toxin (RTX) exotoxin unique to A. pleuropneumoniae and produced by all serovars. The objective of this study was to describe ApxIV antibody responses in serum and oral fluid of pigs.Material and Methods: Four groups of pigs (six pigs per group) were inoculated with A. pleuropneumoniae serovars 1, 5, 7, or 12. Weekly serum samples and daily oral fluid samples were collected from individual pigs for 56 days post inoculation (DPI) and tested by LPS and ApxIV ELISAs. The ApxIV ELISA was run in three formats to detect immunlgobulins M, G, and A (IgM, IgG and IgA) while the LPS ELISA detected only IgG.Results: All pigs inoculated with A. pleuropneumoniae serovars 1 and 7 were LPS ELISA serum antibody positive from DPI 14 to 56. A transient and weak LPS ELISA antibody response was observed in pigs inoculated with serovar 5 and a single antibody positive pig was observed in serovar 12 at ≥35 DPI. Notably, ApxIV serum and oral fluid antibody responses in pig inoculated with serovars 1 and 7 reflected the patterns observed for LPS antibody, albeit with a 14 to 21 day delay.Conclusion: This work suggests that ELISAs based on ApxIV antibody detection in oral fluid samples could be effective in population monitoring for A. pleuropneumoniae.

Introduction: A novel to Europe Schmallenberg virus (SBV) causes clinical disease manifested by reproduction disorders in farm ruminants. In free-living ruminants, SBV antibodies as well as the virus were detected. Recent studies also revealed SBV antibodies in wild boars. The study investigates SBV antibodies occurring in wild boars in Poland at the peak of recent virus epidemics in the country.

Introduction: A novel to Europe Schmallenberg virus (SBV) causes clinical disease manifested by reproduction disorders in farm ruminants. In free-living ruminants, SBV antibodies as well as the virus were detected. Recent studies also revealed SBV antibodies in wild boars. The study investigates SBV antibodies occurring in wild boars in Poland at the peak of recent virus epidemics in the country.Material and Methods: Samples collected from 203 wild boars culled during the 2012/2013 and 2013/2014 hunting season were serologically tested using multi-species cELISA. Attempted neutralisation tests failed due to poor serum quality. RT-PCR was implemented in seropositive and doubtful animals.Results: Two samples collected from wild boar in the winter of 2013 gave a positive result in ELISA, while another two from the 2012/2013 hunting season were doubtful. No SBV RNA was detected in spleen and liver tissues.Conclusion: Low SBV seroprevalence in wild boars, despite high incidence of SBV infections occurring simultaneously in wild ruminants, suggests that boars are unlikely to be a significant reservoir of the virus in the sylvatic environment in Poland.

Introduction: The aim of the study was to analyse selected densitometric and geometric parameters in the third metacarpal bone along the long axis in horses. The densitometric parameters included the cortical and trabecular bone mineral density, while the geometric parameters included the cortical, trabecular, and total areas, strength strain index X, strength strain index Y, and the polar strength strain index.

Introduction: The aim of the study was to analyse selected densitometric and geometric parameters in the third metacarpal bone along the long axis in horses. The densitometric parameters included the cortical and trabecular bone mineral density, while the geometric parameters included the cortical, trabecular, and total areas, strength strain index X, strength strain index Y, and the polar strength strain index. Material and Methods: The parameters were analysed using eight sections from 10% to 80% of the length of the bone. Peripheral quantitative computed tomography was used in the study. Statistical analysis was carried out using the Friedman analysis of variance and post-hoc tests. Results: The proximal metaphyseal region showed the highest predicted resistance to bone fractures in the transverse (back-front) plane, the distal metaphyseal region had the highest predicted resistance to transverse and torsional fractures in the transverse (side-side) plane. The cross-sectional area and the shape of the cross-section of the cortical bone of the MCIII had the highest coefficient of variation. The density of the cortical bone was least variable. Conclusions: The cortical area and cortical bone mineral density assumed the highest values in the diaphyseal region, while the highest total area, trabecular area and trabecular bone mineral density values were obtained in the metaphyseal proximal and distal region.

Introduction: The study aimed to clarify the changes in the concentration of inflammatory mediators, proteases, and cartilage degradation biomarkers in the synovial fluid of joints in an equine osteoarthritis model.

Introduction: The study aimed to clarify the changes in the concentration of inflammatory mediators, proteases, and cartilage degradation biomarkers in the synovial fluid of joints in an equine osteoarthritis model. Material and Methods: Osteoarthritis was induced in eight Mongolian horses by a sterile intra-articular injection of amphotericin B, which was injected into the left carpal joint in a dose of 2 mL (25 mg/mL). The control group comprised five horses which were injected with an equal dose of sterile physiological saline into the left carpal joint. Synovial fluid was obtained at baseline and every week after injection. Test methods were based on ELISA. Results: In the course of the osteoarthritis, the concentration of biomarkers in joint synovial fluid showed an increasing trend. IL-1, IL-6, MMP-9, MMP-13, ADAMTS-5, CS846, GAG, HA, CTX-II, and COMP concentrations sharply increased before the onset of significant symptoms of lameness, whereas TNF-α, MMP-2, and MMP-3 concentrations rose sharply after the occurrence of such symptoms. Conclusion: The results obtained confirm that the concentrations of IL-1, IL-6, MMP-9, MMP-13, ADAMTS-5, CS846, GAG, HA, CTX-II and COMP increase substantially in equine osteoarthritis, which provides a theoretical basis for the rapid diagnosis of the disease.

Introduction: Intentional and accidental poisoning of animals is often caused by readily available commercial pesticides, such as the molluscicide metaldehyde. A retrospective analysis of suspected metaldehyde poisonings between 2014 and 2016 in Italy was conducted. Material and Methods: Biological matrices were collected for toxicological analyses in the course of routine Institute activity. A total of 183 organs from dogs and cats and 49 pieces of bait, here specificall y poisoned food used to lure animals, were analysed and the presence of metaldehyde was confirmed by gas chromatography coupled to mass spectrometry (GC/MS). Results: A high content of metaldehyde was demonstrated in the analysed samples from dogs and cats: 50 (27.3%) animals were found positive for metaldehyde intoxication together with 17 pieces of bait. Conclusion: The study emphasised the need for the control of metaldehyde use by the veterinary service.

Introduction: Intentional and accidental poisoning of animals is often caused by readily available commercial pesticides, such as the molluscicide metaldehyde. A retrospective analysis of suspected metaldehyde poisonings between 2014 and 2016 in Italy was conducted. Material and Methods: Biological matrices were collected for toxicological analyses in the course of routine Institute activity. A total of 183 organs from dogs and cats and 49 pieces of bait, here specificall y poisoned food used to lure animals, were analysed and the presence of metaldehyde was confirmed by gas chromatography coupled to mass spectrometry (GC/MS). Results: A high content of metaldehyde was demonstrated in the analysed samples from dogs and cats: 50 (27.3%) animals were found positive for metaldehyde intoxication together with 17 pieces of bait. Conclusion: The study emphasised the need for the control of metaldehyde use by the veterinary service.

Introduction: Foot quality is essential to the horse’s movement. The barefoot approach favours the animal’s welfare. Environment and selection determine hoof characteristics. Material and Methods: Hoof characteristics of eight Anglo-Arabian (AA) and nine Haflinger (HA) horses were studied. After a preliminary visual analysis of feet, nail samples were collected after trimming for physico-chemical analysis. The parameters were submitted to analysis of variance. A principal component analysis and a Pearson correlation were used to compare mineral contents. Results: The hooves of both breeds were healthy and solid. The hooves of HA horses were longer than those of AA horses (14.90 ±0.30 cm vs 13.10 ±0.60 cm), while the AA hoof was harder than the HA hoof both in the wall (74.55 ±2.95 H vs 60.18 ±2.67 H) and sole (67.00 ±5.87 H vs 43.0 ±4.76 H). In comparison with the sole, the AA hoof wall also had a lower moisture percentage (12.56 ±0.67% vs 20.64 ±0.76%), while crude protein and ash contents were similar in both regions. The AA hoof showed a higher Se content, while the HA hoof had a higher level of macroelements. The negative correlations of K with Cu, Fe, Ni, Pb, and Zn in the AA hoof may indicate osmoregulation activity. Conclusion: The hoof morphology of AA and HA horses met the literature parameters for mesomorphic horses. Both breeds had healthy and well-conformed hooves, useful for sport and recreation activities.

Introduction: Foot quality is essential to the horse’s movement. The barefoot approach favours the animal’s welfare. Environment and selection determine hoof characteristics. Material and Methods: Hoof characteristics of eight Anglo-Arabian (AA) and nine Haflinger (HA) horses were studied. After a preliminary visual analysis of feet, nail samples were collected after trimming for physico-chemical analysis. The parameters were submitted to analysis of variance. A principal component analysis and a Pearson correlation were used to compare mineral contents. Results: The hooves of both breeds were healthy and solid. The hooves of HA horses were longer than those of AA horses (14.90 ±0.30 cm vs 13.10 ±0.60 cm), while the AA hoof was harder than the HA hoof both in the wall (74.55 ±2.95 H vs 60.18 ±2.67 H) and sole (67.00 ±5.87 H vs 43.0 ±4.76 H). In comparison with the sole, the AA hoof wall also had a lower moisture percentage (12.56 ±0.67% vs 20.64 ±0.76%), while crude protein and ash contents were similar in both regions. The AA hoof showed a higher Se content, while the HA hoof had a higher level of macroelements. The negative correlations of K with Cu, Fe, Ni, Pb, and Zn in the AA hoof may indicate osmoregulation activity. Conclusion: The hoof morphology of AA and HA horses met the literature parameters for mesomorphic horses. Both breeds had healthy and well-conformed hooves, useful for sport and recreation activities.

Introduction: The paper presents the method of simultaneous determination of 10 illegal azo dyes in feed by ultra-high performance liquid chromatography coupled with tandem mass spectrometry technique. Material and Methods: The dyes were extracted with hexane, evaporated to dryness, and analysed. Separation was achieved in 7 min in a gradient elution using acetonitrile (A) and 0.1% formic acid (B) as a mobile phase. Results: The validation results showed the repeatability of the method, which was evaluated at three levels (50, 500, and 5,000 μg/kg). All the matrix calibration curves for the working ranges were linear (R² 0.9904 to 1.0), the repeatability was between 2.1% and 24%, and recoveries ranged from 77.9% to 120%. The LOD and LOQ were at 1-2 and 5-10 μg/kg for different dyes, respectively. Furthermore, the method was applied in the homogeneity tests of the in-house prepared feed containing Sudan I at the levels of 0.5, 5, and 50 mg/kg. Conclusions: A sensitive, selective, and fast multiresidue method was successfully developed and validated. Its robustness was confirmed by the analysis of an experimental feed containing Sudan I.

Introduction: The paper presents the method of simultaneous determination of 10 illegal azo dyes in feed by ultra-high performance liquid chromatography coupled with tandem mass spectrometry technique. Material and Methods: The dyes were extracted with hexane, evaporated to dryness, and analysed. Separation was achieved in 7 min in a gradient elution using acetonitrile (A) and 0.1% formic acid (B) as a mobile phase. Results: The validation results showed the repeatability of the method, which was evaluated at three levels (50, 500, and 5,000 μg/kg). All the matrix calibration curves for the working ranges were linear (R2 0.9904 to 1.0), the repeatability was between 2.1% and 24%, and recoveries ranged from 77.9% to 120%. The LOD and LOQ were at 1-2 and 5-10 μg/kg for different dyes, respectively. Furthermore, the method was applied in the homogeneity tests of the in-house prepared feed containing Sudan I at the levels of 0.5, 5, and 50 mg/kg. Conclusions: A sensitive, selective, and fast multiresidue method was successfully developed and validated. Its robustness was confirmed by the analysis of an experimental feed containing Sudan I.

Introduction: Pregnancy is a physiological state in which the immune system undergoes certain changes. On the one hand, by depleting cell defence mechanisms, it favours development and maintenance of the pregnancy. At the same time cells of the immune system ensure resistance to many risk factors, including infectious agents.Material and Methods: The study was carried out on 24 Polish Konik breed mares which were divided into two equal groups. The first group (group I) included mares living in the reserve. The second group (group II) comprised mares maintained under conventional conditions in the stables. The blood samples were collected for the first time in the perinatal period, i.e. 2 weeks before parturition (trial 0), then within the first 24 h after delivery, and then on 7ᵗʰ and 21ˢᵗ day after foaling. Flow cytometric analysis of lymphocyte expressing TCD4+, TCD8+, CD2+, and MHC class II antigens was performed.Results: Before the delivery, in group I there was a significantly higher CD4:CD8 ratio compared to group II (P ≤0.05). Similarly, significantly increased CD4:CD8 ratio in group I was noted within 24 h after parturition (P ≤0.001) and it was also observed on 7ᵗʰ day (P ≤0.03) and 21ˢᵗ day after foaling (P ≤0.02). In the first 24 h after parturition, a significant decline of lymphocytes CD8+ (P ≤0.02) was noted. No significant differences in terms of lymphocytes CD2+ and CD3+ were observed. Expression of MHC-II molecules before and after the parturition was higher in group I compared to group II; however, the difference between the groups was not significant.Conclusion: The results obtained indicate that mares living in the reserve display higher activity of cell defence mechanisms.

Introduction: Pregnancy is a physiological state in which the immune system undergoes certain changes. On the one hand, by depleting cell defence mechanisms, it favours development and maintenance of the pregnancy. At the same time cells of the immune system ensure resistance to many risk factors, including infectious agents.

Introduction: The transition period is the most challenging time for dairy cattle, which is characterised not only by negative energy balance but also by fatty tissue mobilisation.Material and Methods: The efficiency of energy pathways, β-oxidation in WBC and glycolysis in RBC (based on deoxyglucose transmembrane transport) were estimated. Insulin in blood plasma was determined using ELISA.Results: After calving and up to one month after delivery, a significant drop in blood plasma level was noticed, simultaneously with a rise in β-oxidation from 18.93 ±3.64 to 30.32 ±5.28 pmol/min/mg protein in WBC. A strong negative correlation between these two indices (r = −0.68) was found. During the period of transition to lactation an increase in glucose cross-membrane transportation from 41.44 ±4.92 to 50.49 ±6.41 μmol/h/g Hb was observed. A strong positive correlation between glucose transportation in RBC and β-oxidation in WBC (r = 0.71) was noticed. These data are in agreement with results of studies on dairy cows using liver slices from dairy cows in late pregnancy and different stages of lactation, in which changes in gene expression were analysed.Conclusion: It seems that measuring fatty acids oxidation and glycolysis using isolated blood cells may be an adequate and relatively simple method for energy state analysis to estimate the state of dairy cow metabolism and animal health.

Introduction: The transition period is the most challenging time for dairy cattle, which is characterised not only by negative energy balance but also by fatty tissue mobilisation.