Evaluation of oscillometric blood pressure monitor BLT M9000 VET in anaesthetised healthy adult dogs.

To examine agreement in anaesthetised dogs between invasive blood pressure measurements and measurements obtained with an oscillometric blood pressure monitor. MATERIALS AND METHODS: Paired invasive and oscillometric measurements were taken in 24 dogs every 5 minutes during anaesthesia (9 to 37 measurements per dog). Agreement between measurement methods was explored using Bland-Altman plots. To determine the accuracy of the oscillometric measurements, the results were compared with the guidelines recommended by the American College of Veterinary Internal Medicine and the Association for the Advancement of Medical Instrumentation. RESULTS: In total, 493 paired readings were obtained: 98·6% of oscillometric readings were successful. Biases (±standard deviation) for oscillometric readings of systolic arterial blood pressure and mean arterial blood pressure were 2·1 (±11·5) and -9·8 (±7·6) mm Hg, indicating slight over- and under-estimation, respectively, versus invasive measurements. More than 50% and 80% of systolic arterial and mean arterial pressure measurements were within 10 and 20 mmHg of invasively measured values, respectively. A large negative bias (-14·1 ±9·2 mmHg) against invasive measurements revealed that the oscillometric measures of diastolic arterial blood pressure measurements were under-estimated. CLINICAL SIGNIFICANCE: In healthy adult anaesthetised dogs, this oscillometric monitor met the American College of Veterinary Internal Medicine requirements for systolic arterial and mean arterial pressure measurement but failed to meet the requirements for the measurement of diastolic arterial pressure.

Improvement of liquid stored boar semen quality by removing low molecular weight proteins and supplementation with α-tocopherol.

Seminal plasma contains low-molecular weight components that can exert a harmful effect on sperm function. We have evaluated the effects of removing low-molecular weight components from seminal plasma and adding α-tocopherol on boar semen quality after 72h of liquid storage. Semen was evaluated on the basis of motility, morphology, acrosome integrity, plasma membrane modifications, mitochondrial activity, DNA fragmentation and lipid peroxidation. Thiobarbituric acid reactive substances (TBARS), 8-isoprostane, and antioxidant status (total antioxidant capacity (TAC) and superoxide dismutase activity (SOD)) were measured in seminal plasma. Removal of low-molecular weight components from seminal plasma, together with the addition of α-tocopherol, kept the lipid peroxidation and mitochondrial activity and DNA fragmentation at the same level as in native semen samples. Dialysing semen and adding 200µM of α-tocopherol led to higher progressive motility, a higher proportion of morphologically normal spermatozoa and a significantly lower level of acrosomal reacted spermatozoa compared to non-dialyzed semen samples after 72h of storage. In conclusion, liquid stored boar semen was better preserved, and oxidative stress in the semen was reduced when semen was dialyzed and α-tocopherol was added prior to storage.

The Effect of Seasonal Thermal Stress on Lipid Mobilisation, Antioxidant Status and Reproductive Performance in Dairy Cows.

Heat stress is a major factor contributing to low fertility of dairy cows with a great economic impact in dairy industry. Heat-stressed dairy cows usually have reduced nutrient intake, resulting in a higher degree of negative energy balance (NEB). The aim of this study was to investigate the seasonal thermal effect on lipid metabolism, antioxidant activity and reproductive performance in dairy cows. Thirty-two healthy dairy heifers were included in the study. According to the ambient temperature, animals were divided into two groups: winter (N = 14) and summer season (N = 18). Metabolic parameters, paraoxonase-1 (PON1) activity and total antioxidant status (TAS) were monitored at the time of insemination (basal values) and from 1 week before until 8 weeks after calving. Number of services per conception and calving-to-conception (CC) interval were calculated from the farm recording data. Serum triglyceride, non-esterified fatty acids (NEFA) and beta-hydroxybutyrate (BHB) concentrations were significantly increased after calving in summer compared to winter, indicating higher degree of NEB in cows during summer. PON1 activity was significantly decreased after calving in both summer and winter group. TAS concentration was significantly lower in summer than that in winter. A significantly higher number of services were needed for conception in summer compared to winter, and CC interval was significantly longer in summer than that in winter as well. Additionally, reproductive performance significantly correlated with the severity of NEB, suggesting that lipid mobilization and lower antioxidant status contributed to poor reproduction ability in dairy cows during hot months.

Manageable risk factors associated with bacterial and coliform counts in unpasteurized bulk milk in Flemish dairy herds.

Associations between herd management practices and both bacterial counts (BC) and coliform counts (CC) from 254 and 242 dairy herds in Flanders (Belgium), respectively, were studied. Data were analyzed using multivariable, multilevel linear regression analysis, allowing variance components analyses. Both BC and CC fluctuated throughout the year, although the milk quality parameters followed an opposite pattern. Bacterial count values decreased with each increase of the cleaning frequency of the cubicles (once per week, once per day, twice per day, or more than twice per day) between January and March. Herds with a conventional milking parlor had substantially lower BC than herds where the cows were milked using an automatic milking system. Lower BC were observed when the milking parlor was equipped with an automatic cluster removal system, when premilking teat disinfection was applied, when the dry cows were supplemented with a mix of minerals and vitamins, and when the teats were prepared either first wet and dried or via an automatic milking system. Milking cows with a high-pipeline milking parlor setup or with an automatic milking system was associated with substantially higher CC values. Herds where prepartum heifers were often treated with antimicrobials before calving had a lower CC than farms where heifers were either not or only rarely treated. Most variation in BC and CC resided at the herd level rather than at the observation level, indicating that management is important in the control of both BC and CC. Still, only a small proportion of the total variance was explained by factors capturing information related to the milking, herd health, and dry cow management, which suggests that the bacteriological milk quality and, in particular, CC is primarily driven by other factors than the ones included in this study.

Lipid mobilisation and oxidative stress as metabolic adaptation processes in dairy heifers during transition period.

The objective of this study was to evaluate metabolic disorders and oxidative stress in dairy heifers during the transition period. Possible relationships between lipid mobilisation indicators and oxidative stress markers were investigated as well. Nineteen dairy heifers were included in the study. Blood samples were collected at the time of estrus synchronisation in heifers, at insemination, three weeks after insemination, one week before calving, at calving and 1, 2, 4 and 8 weeks postpartum. Common metabolic parameters, beta-hydroxybutyrate (BHB), free fatty acids (FFA), paraoxonase-1 (PON1) activity and total antioxidative status (TAS) were analysed. Around insemination, no significant difference was observed in the majority of tested parameters (P>0.05). However, the transition period markedly affected the concentration of triglycerides, total cholesterol, HDL-C, BHB, FFA, TAS and PON1activity. Positive correlations between PON1 activity and total cholesterol, HDL-C and triglycerides were noted but inverse correlations with FFA, BHB and bilirubin were found indicating that PON1 activity changed with lipid metabolism and was influenced by negative energy balance. These findings suggest that lipid mobilisation and oxidative stress are part of a complex metabolic adaptation to low energy balance which reaches equilibrium later in advanced lactation.

Diagnostic evaluation of fat to protein ratio in prolonged calving to conception interval using receiver operating characteristic analyses.

Evaluation of fat to protein ratio (FPR) in milk was performed for the prediction of calving to conception interval (CC) in dairy cows. Reproduction parameters of 51 high yielding dairy cows were calculated from farm recording data and milk data record from a regular dairy control. Spearman rank correlation analysis was used to determine the correlation between reproductive parameters of the herd and the milk data record. In the time interval of 75-90 days postpartum the highest correlation was found between FPR and CC (r = 0.414, p < 0.05). Receiver operating characteristics (ROC) analysis was used to evaluate the FPR to distinguish between cows with different CC. The optimal cut-off value at FPR of 1.34 provided the best discrimination power according to CC of 120 days. FPR at 1.44 enabled us 91.7% correct identification of cows with CC above 140 days, where the highest area under the ROC curve of 0.759 and likelihood ratio of 7.2 were observed. FPR

Spectrophotometric application of resazurin reduction assay to evaluate boar semen quality.

The resazurin reduction assay depends on the ability of metabolically active cells to reduce the resazurin redox dye to resorufin. In the present study we applied and made a diagnostic evaluation of a spectrophotometric application of the resazurin reduction assay to assess the colour change of resazurin reduction in butanol extracted colour to evaluate boar semen quality. Forty-one samples of boar semen from various breeds were included in the study. The absorption peaks for resazurin and resorufin were found to be 610 and 575 nm, respectively. Absorbance at 610 nm, where the minimum overlap of the two peaks was observed, was used in further analysis. Spearman rank correlation analysis was used to determine the correlation between the resazurin reduction assay and various semen parameters. The highest correlations were observed with the concentration of motile spermatozoa (r = -0.841; p < 0.001), sperm concentration (r = -0.833; p < 0.001), sperm index (-0.826; p < 0.001) and concentration of viable spermatozoa (r = -0.763; p < 0.001). Sensitivity and specificity, at 94.1 and 91.7%, respectively, indicate that the present test is highly accurate in discriminating between the samples according to the sperm index. When motile sperm concentration was used to distinguish between good and poor samples, high sensitivity (93.6%) was also found, whereas the test was only moderately, 80%, specific. The stability of butanol extracts in terms of A610 at different times of measurement confirmed that the resazurin reduction could be spectrophotometrically measured within 7 days from the time of assay performance, making the assay much more useful. Based on these results, the assay could be used as an additional tool for evaluating the quality of boar semen.

Serum-specific antibodies in rabbits naturally infected with Trichophyton mentagrophytes.

In the present study, we investigated the humoral immune response of rabbits to Trichophyton mentagrophytes (sensu lato) proteins obtained from keratin-rich media in vitro. The test rabbits were naturally infected with T. mentagrophytes. The production of keratinolytic enzymes in T. mentagrophytes was stimulated by growing cultures with keratin as a sole nitrogen source. The proteins were isolated from a protein extract prepared from the fungal mat. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) showed three bands. Bands with Mr 20 and 30 kDa were glycosylated, whereas a band of 18 kDa was not. The rabbits' humoral immune responses to the total protein extract of T. mentagrophytes and to the proteins with keratinolytic activity was studied by immunoblotting. IgG from infected rabbits' sera revealed eight dominant bands with apparent molecular weights between 20 and 75 kDa. Bands of 20, 30 and 33 kDa appeared with a frequency rate of 76% only on immunoblots of infected rabbit serum. Using an indirect enzyme-linked immunosorbent assay (ELISA), we observed a significant increase in specific antibodies in a group of infected rabbits compared to a control group (P < 0.001). The ELISA exhibited 95% sensitivity and 83% specificity at the optimal cut-off value, with 90% predictive values of a positive and a negative result. Under these conditions, it could be used in the accurate detection of specific antibodies in sera of infected rabbits.

Detection by ELISA of the humoral immune response in rabbits naturally infected with Trichophyton mentagrophytes.

An indirect enzyme linked immunosorbent assay (ELISA) was developed and its diagnostic potential evaluated for rabbits infected by Trichophyton mentagrophytes. Within-run and between-run coefficient of variance varied from 2.3 to 7.7% and from 5.9 to 8.5%, respectively, indicating satisfactory reproducibility of the ELISA. There was no significant cross-reaction with antigens of Microsporum canis, Malassezia pachydermatis and Aspergillus fumigatus. The level of specific IgG to Trichophyton mentagrophytes was measured in sera of 25 11-week-old and 12 younger infected rabbits. There was no significant difference in the IgG level between 12 5-week-old infected rabbits and controls (p = 0.38). The antibody response was higher in 12 7-week-old rabbits compared with controls (p = 0.001). The IgG level in 25 11-week-old rabbits differed from the controls very significantly (p < 0.0001). Increased specific IgG in 11-week-old rabbits exhibited 96% sensitivity and 94% specificity. Predictive values of a positive and a negative test were 96 and 94%, respectively. Western immunoblotting associated three protein bands (21.5, 31, 44 kDa) with Trichophyton mentagrophytes infection.