The plate counting method widely used at present to discern viable from non-viable Brucella in the host or cell is time-consuming and laborious. Therefore, it is necessary to establish a rapid, simple method for detecting and counting viable Brucella organisms. Using propidium monoazide (PMA) to inhibit amplification of DNA from dead Brucella, a novel, rapid PMA-quantitative PCR (PMA-qPCR) detection method for counting viable Brucella was established. The standard recombinant plasmid with the target BCSP31 gene fragment inserted was constructed for drawing a standard curve. The reaction conditions were optimised, and the sensitivity, specificity, and repeatability were analysed. The optimal exposure time and working concentration of PMA were 10 min and 15 μg/mL, respectively. The correlation coefficient (R²) of the standard curve was 0.999. The sensitivity of the method was 10³ CFU/mL, moreover, its specificity and repeatability also met the requirements. The concentration of B. suis measured by the PMA-qPCR did not differ significantly from that measured by the plate counting method, and the concentrations of viable bacteria in infected cells determined by the two methods were of the same order of magnitude. In this study, a rapid and simple PMA-qPCR counting method for viable Brucella was established, which will facilitate related research.

Introduction: Aujeszky’s disease virus (ADV) infects a wide range of animals, including members of the Suidae family, i.e. domestic and wild pigs, as well as wild boar. Since wild boar are a potential ADV reservoir and a source of infection for domestic pigs, the aim of the study was to evaluate ADV antibody prevalence in the Polish wild boar population, during the years 2011 to 2014. Material and Methods: Wild boar blood samples were collected during three consecutive hunting seasons; i.e. 2011/2012, 2012/2013, and 2013/2014, and tested for ADV antibodies by ELISA. Results: ADV antibodies were detected in samples from all tested voivodships. The average seroprevalence reached 32.2%. Seroprevalence, over the examined hunting seasons, was 27.4% in 2011/2012, 32.4% in 2012/2013, and 35.5% in 2013/2014. The highest percentage of seroreagents was detected in four voivodships, situated along the western border of Poland, i.e. Zachodnio-Pomorskie (ZP), Lubuskie (LB), Dolnośląskie (DS), and Opolskie (OP). This area is positively correlated with the highest density of the wild boar population and the highest wild boar hunting bag. Conclusion: The results of this study confirm that the wild boar population may still pose a threat to domestic pigs, which is of special importance at the final stage of Aujeszky’s disease eradication programme in Poland.

Introduction: The purpose of the current study was to evaluate the blood glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) levels under seasonal variations in dairy cows during transition period, and to assess the relationship between chosen reproductive parameters, GSH-Px, and MDA. Material and Methods: Holstein cows calving in January were assigned into winter group (n = 42), while cows calving in August were assigned into summer group (n = 42). Blood samples were collected from the jugular vein 21, 14, and 7 days before calving, at calving (0 day), and 7, 14, and 21 days after calving. Reproductive parameters obtained from farm records were evaluated. Results: In both groups of cows, GSH-Px activity decreased from 21 days before calving to day 0, and it gradually continued to increase until 21 days after calving. GSH-Px activity was higher in winter group compared to summer group during the transition period (P < 0.05). MDA levels in both groups increased over time starting from 21 days before calving to 0 day, but it gradually decreased thereafter. MDA levels were higher in summer group compared to winter group during the transition periods (P < 0.05). Summer group of cows showed higher intervals of calving-to-oestrus, calving-to-conception, and higher insemination index (P < 0.01). Negative correlation was recorded between GSH-Px and MDA during all examination days (P < 0.01). MDA levels correlated with calving to conception interval on day 21 before calving and day 0 (P < 0.01) and insemination index on day 0 and 21 days after calving (P < 0.01). GSH-Px activity was negatively correlated with calving to conception interval on day 21 before calving, day 0, and 21 days (P < 0.01) after calving. Negative correlation on day 21 before calving and day 0 was also determined between GSH-Px and insemination index (P < 0.01). Conclusion: This study showed that blood oxidant and antioxidant levels have affected the fertility parameters in cows under seasonal variations.

Introduction: The predictive value of selected parameters in the risk of ketosis and fatty liver in dairy cows was determined. Material and Methods: In total, 21 control and 17 ketotic Holstein Friesian cows with a β-hydroxybutyrate (BHBA) concentration of 1.20 mmol/L as a cut-off point were selected. The risk prediction thresholds for ketosis were determined by receiver operating characteristic (ROC) curve analysis. Results: In the ketosis group, paraoxonase-1 (PON-1) activity and concentration of PON-1 and glucose (GLU) were decreased, and aminotransferase (AST) activity as well as BHBA and non-esterified fatty acid (NEFA) contents were increased. The plasma activity and concentration of PON-1 were significantly positively correlated with the level of plasma GLU. The plasma activity and concentration of PON-1 were significantly negatively correlated with the levels of AST and BHBA. According to ROC curve analysis, warning indexes of ketosis were: plasma PON-1 concentration of 46.79 nmol/L, GLU concentration of 3.04 mmol/L, AST concentration of 100 U/L, and NEFA concentration of 0.82 mmol/L. Conclusion: This study showed that the levels of PON-1, GLU, AST, and NEFA could be used as indicators to predict the risk of ketosis in dairy cows.

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: The aim of the study was to investigate changes in the serum levels of adiponectin and TNF-α, as well as insulin sensitivity, and to elucidate the possible relationship among the parameters and negative energy balance during the periparturient period of dairy cows.Material and Methods: Thirty primiparous Holstein dairy cows were selected for the study. Blood samples were collected from each cow seven days before the expected calving date, on the calving day, and 7, 14, and 21 days after calving. Blood non-esterified fatty acids (NEFA), β-hydroxybutyric acid (BHBA), glucose, insulin, adiponectin, and TNF- α levels were measured. Revised Quantitative Insulin Sensitivity Check Index (rQUICKI) was calculated using data on NEFA, insulin, and glucose concentrations.Results: When compared to prepartum levels, serum concentration of adiponectin significantly increased on day 21 postpartum. The rQUICKI increased and NEFA levels decreased on day 7 after parturition. Insulin and glucose levels decreased on days 7, 14, and 21 postpartum when compared with prepartum levels. BHBA levels decreased on day 21 and TNF- α concentration also decreased on days 7, 14, and 21 postpartum. Adiponectin levels positively correlated with NEFA during the preparturient period. Negative correlation was detected between adiponectin and rQUICKI on calving day and on 14ᵗʰ day after parturition. TNF- α concentration positively correlated with glucose levels on day 7 prepartum and on 21ˢᵗ day postpartum and with rQUICKI on 21ˢᵗ day postpartum. Negative correlation was detected between adiponectin level and insulin sensitivity.Conclusion: Based on the results of the study, we concluded that adiponectin could possibly increase insulin sensitivity when blood NEFA concentrations are elevated.

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.

OBJECTIVE To measure serum homocysteine concentrations in dogs with myxomatous mitral valve disease (MMVD) and identify any association between this variable and stage of MMVD. ANIMALS 53 client-owned dogs with MMVD and 10 healthy control Beagles. PROCEDURES Dogs with MMVD were allocated to 3 groups in accordance with the staging system for chronic valvular heart disease in dogs and cats of the American College of Veterinary Internal Medicine. Blood samples were collected from all dogs, and serum homocysteine and cardiac troponin 1 concentrations were measured by enzyme immunoassay and chemiluminescence immunoassay, respectively. Analyte values were tested for associations with each other and with stage of MMVD. RESULTS A significant correlation was identified between serum homocysteine concentration and stage of MMVD. Mean ± SD concentrations were 6.72 ± 1.65 μmol/L for control dogs, 13.37 ± 4.16 μmol/L for dogs with stage B MMVD, 18.86 ± 6.73 μmol/L for dogs with stage C disease, and 28.26 ± 4.48 μmol/L for dogs with stage D disease. In addition, serum homocysteine concentration was correlated with serum cardiac troponin 1 (r = 0.34) and creatinine (r = 0.46) concentrations, systolic blood pressure (r = 0.57), and left atrium-to-aortic root ratio (r = 0.28), all of which were positively correlated with stage of MMVD. CONCLUSIONS AND CLINICAL RELEVANCE Serum homocysteine concentrations of dogs with MMVD were significantly higher than those of control dogs, and significant correlations were identified between these values and several risk factors for heart failure. Measurement of serum homocysteine concentration may be useful in the prediction of severity of disease in dogs with MMVD.

OBJECTIVE To evaluate accuracy and reliability of 3 novel goniometers for measurement of canine stifle joint angles and compare the results with those obtained with a universal goniometer (UG). SAMPLE 8 pelvic limbs from 4 canine cadavers. PROCEDURES Each limb was secured to a wooden platform at 3 arbitrarily selected fixed stifle joint angles. Goniometry was performed with 2 smartphone-based applications (novel goniometers A and B), a digital goniometer (novel goniometer C), and a UG; 3 evaluators performed measurements in triplicate for each angle with each device. Results were compared with stifle joint angle measurements on radiographs (used as a gold standard). Accuracy was determined by calculation of bias and total error, coefficients of variation were calculated to estimate reliability, and strength of linear association between radiographic and goniometer measurements was assessed by calculation of correlation coefficients. RESULTS Mean coefficient of variation was lowest for the UG (4.88%), followed by novel goniometers B (7.37%), A (7.57%), and C (12.71%). Correlation with radiographic measurements was highest for the UG (r = 0.97), followed by novel goniometers B (0.93), A (0.90), and C (0.78). Constant bias was present for all devices except novel goniometer B. The UG and novel goniometer A had positive constant bias; novel goniometer C had negative constant bias. Total error at 50° and 100° angles was > 5% for all devices. CONCLUSIONS AND CLINICAL RELEVANCE None of the devices accurately represented radiographically measured stifle joint angles. Additional veterinary studies are indicated prior to the use of novel goniometers in dogs.

OBJECTIVE To evaluate the correlation coefficient of the ratio between diameter of the caudal vena cava (CVC) and diameter of the aorta (Ao) in dogs as determined ultrasonographically with systolic pressure variation (SPV). ANIMALS 14 client-owned dogs (9 females and 5 males; mean ± SD age, 73 ± 40 months; mean body weight, 22 ± 7 kg) that underwent anesthesia for repair of skin wounds. PROCEDURES Anesthesia was induced. Controlled mechanical ventilation with a peak inspiratory pressure of 8 cm H2O was immediately started, and SPV was measured. During a brief period of suspension of ventilation, CVC-to-Ao ratio was measured on a transverse right-lateral intercostal ultrasonographic image obtained at the level of the porta hepatis. When the SPV was ≥ 4 mm Hg, at least 1 bolus (3 to 4 mL/kg) of Hartmann solution was administered IV during a 1-minute period. Bolus administration was stopped and the CVC-to-Ao ratio measured when SPV was < 4 mm Hg. Correlation coefficient analysis was performed. RESULTS 28 measurements were obtained. The correlation coefficient was 0.86 (95% confidence interval, 0.72 to 0.93). Mean ± SD SPV and CVC-to-Ao ratio before bolus administration were 7 ± 2 mm Hg and 0.52 ± 0.16, respectively. Mean ± SD SPV and CVC-to-Ao ratio after bolus administration were 2 ± 0.6 mm Hg and 0.91 ± 0.13, respectively. CONCLUSIONS AND CLINICAL RELEVANCE In this study, the CVC-to-Ao ratio was a feasible, noninvasive ultrasonographically determined value that correlated well with SPV.