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: The aim of this study was to evaluate the effect of lipoprotein fraction isolated from ostrich egg yolk (LPFo) on the metabolic activity of boar spermatozoa following liquid semen storage in different extenders and temperatures. Material and Methods: Boar ejaculates were extended in Androhep, Beltsville thawing solution (BTS), and Martín-Rillo and Alias (MR-A) without (control) or with the addition of LPFo and stored for three days at either 5°C or 16°C. The analysed sperm parameters included total motility (TMOT), plasma membrane integrity (PMI), mitochondrial membrane potential (MMP), oxygen consumption, and adenosine triphosphate (ATP) production. Results: The sperm metabolic activity seemed to be higher in the LPFo-based extenders following storage for three days, irrespective of the storage temperature. Compared with the LPFo-free extenders, significantly higher (P < 0.05) sperm PMI and MMP were observed in BTS and MR-A extenders supplemented with LPFo during storage for three days at 5°C. Spermatozoa stored in the BTS-LPFo extender exhibited higher (P < 0.05) TMOT and oxygen consumption, whereas higher (P < 0.05) PMI was observed in spermatozoa stored in Androhep-LPFo and MR-A-LPFo for three days at 16°C. No significant differences (P > 0.05) in ATP content were observed between the LPFo-free and LPFo-based extenders during storage. Conclusions: Supplementation of LPFo to semen extenders had varying effects on the metabolic activity of boar spermatozoa stored at different temperatures. It can be suggested that the interactions of various components of the extenders and seminal plasma with LPFo exert beneficial effects on the sperm metabolic activity during liquid storage of boar semen.

Introduction: The study aimed at evaluating oxidative stress using malondialdehyde (MDA), total antioxidant capacity (TAC), total oxidant status (TOS), and oxidative stress index (OSI) markers in sheep naturally infected with Psoroptes ovis (Acari).Material and Methods: The study was performed on 40 sheep divided into two equal groups: a healthy group (group I) and a group naturally infected with Psoroptes ovis (group II). The sera were obtained by centrifuging blood samples collected from the vena jugularis and serum MDA level changes in the samples were measured spectrophotometrically. Commercially available test kits were used for the measurement of TAC and TOS levels. The percentage ratio of TOS level to TAC level was accepted as OSI.Results: The serum malondialdehyde, total oxidant status levels, and oxidative stress index increased significantly (P < 0.01) in group II, while the serum total antioxidant capacity levels decreased significantly (P < 0.01) in this group. Negative correlations between total antioxidant capacity and total oxidant status and total antioxidant capacity and malondialdehyde, and a positive correlation between total oxidant status and malondialdehyde were found in infected sheep.Conclusion: The obtained results indicated the relationship between oxidant/antioxidant imbalance and Psoroptes ovis infection in sheep. Their MDA, TAC, TOS, and OSI markers may be used to determine the oxidative stress in natural infections with Psoroptes ovis.

Introduction: The purpose of the study was to determine and model the growth rates of L. monocytogenes in cooked cured ham stored at various temperatures. Material and Methods: Samples of cured ham were artificially contaminated with a mixture of three L. monocytogenes strains and stored at 3, 6, 9, 12, or 15°C for 16 days. The number of listeriae was determined after 0, 1, 2, 3, 5, 7, 9, 12, 14, and 16 days. A series of decimal dilutions were prepared from each sample and plated onto ALOA agar, after which the plates were incubated at 37°C for 48 h under aerobic conditions. The bacterial counts were logarithmised and analysed statistically. Five repetitions of the experiment were performed. Results: Both storage temperature and time were found to significantly influence the growth rate of listeriae (P < 0.01). The test bacteria growth curves were fitted to three primary models: the Gompertz, Baranyi, and logistic. The mean square error (MSE) and Akaike’s information criterion (AIC) were calculated to evaluate the goodness of fit. It transpired that the logistic model fit the experimental data best. The natural logarithms of L. monocytogenes’ mean growth rates from this model were fitted to two secondary models: the square root and polynomial. Conclusion: Modelling in both secondary types can predict the growth rates of L. monocytogenes in cooked cured ham stored at each studied temperature, but mathematical validation showed the polynomial model to be more accurate.

Introduction: In the European Union the use of steroid growth promoters is prohibited under Council Directive 96/22/EC. For effective control of illegal use of natural steroids, highly sensitive analytical methods are required, because sex hormones can be present in very low concentrations in biological samples. The aim of the study was to develop a confirmatory method for the detection of testosterone in bovine serum at ppt level. Material and Methods: 17β-testosterone and internal standards of 17β-testosterone-d2 were extracted from serum samples with a mixture of tert-butyl methyl ether/petroleum ether and were directly analysed by an LC/MS/MS on QTRAP 5500 instrument with a TurboIon-Spray source operating in a positive ionisation mode. Chromatographic separation was achieved on the analytical column Inertsil® ODS-3 with an isocratic elution using mobile phase consisting of acetonitrile, methanol, and water. Method validation has been carried out in accordance with the Commission Decision 2002/657/EC. Results: The method was characterised by good recovery (82%) and precision (R.S.D 17 %). Decision limit (CCα) and detection capability (CCβ) was 0.05 μg L⁻¹ and 0.09 μg L⁻¹ respectively. The method met the criteria set out in Commission Decision 2002/657/EC for the purpose of confirmation in terms of retention time and ion ratio in the whole range of its application. Conclusions: The developed method is specific and sensitive, suitable for measuring the natural level of testosterone in blood of cattle and for use in routine control programme for the detection of this hormone in bovine serum.

Introduction: There are many veterinary products containing β-lactam antibiotics which are used for mastitis treatment in cows. The aim of the study was to determine whether mastitis could have any effect on amoxicillin (AMX) or penicillin G procaine (PEN) withdrawal period from milk, in the context of current maximum residue limits established by the European Commission. Material and Methods: The study was conducted on 17 dairy Black and White cows with clinical mastitis during the lactation period. The first group (n = 8) received 200 mg of amoxicillin (AMX), whereas the second group (n = 9) received 200,000 IU/mg of penicillin G procaine (PEN) by intramammary administration. For the measurement of AMX and PEN concentrations in milk, the liquid chromatography tandem mass spectrometry method was applied. Pharmacokinetic calculations were performed using Phoenix WinNonlin 6.4 software. Results: The determined AMX and PEN half-life values in the mammary gland suggest that the drug withdrawal is at a level of 99.9% within 81 h (≈3.5 days) and 116 h (≈5 days) after administration of AMX and PEN, respectively. The present research indicates that, at 60 h after administration, the average PEN concentration in the milk from cows with clinical signs of mastitis may still reach 4.96 g/kg and that of AMX can even be 6.92 g/kg. Conclusion: The results obtained confirm that, in mastitis cases, a 72-h withdrawal period is sufficient for elimination of AMX to a lower level than the established maximum residue limit (MRL) values. However, in the case of PEN, at 69 h after administration, the drug concentration may be close to that of the determined MRL.

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: A model of fatty liver in postpartum sheep was established to measure blood paraoxonase 1 (PON1) and other biochemical indicators, which were used to predict fatty liver in sheep. Material and Methods: Sheep were assigned into two experimental groups: a fatty liver group (T, n = 10) and a healthy control group (C, n = 5). PON1 enzyme activity towards paraoxon as a substrate was quantified spectrophotometrically. The results were analysed by t-test and pearson correlation coefficient. Disease was predicted by binary logistic analysis, and diagnostic thresholds were determined by receiver operatingcharacteristic (ROC) analysis. Results: The activity of serum PON1 in group T was significantly decreased (P < 0.05) when compared with C group, and liver lipid content and the levels of serum BHBA, NEFA, and TG were significantly increased (P < 0.05). Thresholds were lower than 74.0 U/mL for PON1, higher than 0.97 mmol/L for β-hydroxybutyrate, higher than 1.29 mmol/L for non-esterified fatty acids, higher than 0.24 mmol/L for triglycerides, and lower than 71.35 g/L for total protein. Conclusion: This study verified that PON1, BHBA, NEFA, TG, and TP could be used to predict the risk of fatty liver in sheep.

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: Infections with bovine foamy virus (BFV) were found in many countries but there is a lack of large-scale surveys on the prevalence of BFV among dairy cattle. The aim of this study was to develop and validate the recombinant Gag protein-based ELISA and to estimate the prevalence of antibodies against BFV. Material and Methods: Gag coding region from BFV was cloned into expression vector pT7Arg-STOP, which expressed a high level of recombinant Gag protein from E.coli. The ELISA was standardised, and the cut-off value and sensitivity and specificity of the test were calculated using a receiver operating characteristic and Bayesian estimation. Results: A total of 3,051 serum samples were tested by ELISA and 939 (30.8%) sera were recognised as positive. When Bayesian approach was used, the overall true BFV prevalence was 29.7% (95% CI: 25.9-33.4%). Conclusion: Expressed Gag protein of BFV has been used successfully as an antigen for ELISA. Eventually, this study provides basic information about the epidemiological status of infection with BFV in dairy cattle in Poland, which can be used for further studies on dissemination and transmission of BFV infection.