Petra Zrimšek

Comparison of high definition oscillometric and Doppler ultrasound devices with invasive blood pressure in anaesthetized dogs.

OBJECTIVE To use the American College of Veterinary Internal Medicine (ACVIM) validation criteria to evaluate the performance of high definition oscillometric (HDO) and Doppler blood pressure measurement techniques against invasive blood pressure measurements in anaesthetized dogs. STUDY DESIGN Prospective clinical study. ANIMALS Twenty client-owned dogs. MATERIALS AND METHODS Invasive blood pressure was measured using a catheter inserted into a pedal artery and an electronic transducer. The sites of cuff placement for the HDO measurements were the mid antebrachium or the proximal tail and, for the Doppler technique, the distal tibia. Agreement between invasive and non-invasive blood pressure measurements was estimated by the Bland-Altman method. RESULTS Only 10% and 34% of Doppler measurements were within 10 and 20 mmHg of invasive blood pressure values, respectively. The Doppler device failed to meet the ACVIM validation criteria for blood pressure measurement devices. The best agreement between HDO and invasive blood pressure measurement technique was observed for mean arterial blood pressure (MAP); 67% and 95% of readings were within 10 and 20 mmHg of invasive blood pressure values respectively. In addition, 52% and 87% of diastolic arterial blood pressure (DAP) measurements were within 10 and 20 mmHg of invasive readings. High definition oscillometric readings did not meet ACVIM recommended limits for SAP. CONCLUSION AND CLINICAL RELEVANCE The Doppler technique overestimated and the HDO device showed limited agreement with invasive blood pressure measurement in anaesthetized dogs. High definition oscillometry met most of the ACVIM requirements for MAP and DAP while the Doppler technique did not.

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.

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.

PLASMA MALONDIALDEHYDE, BIOCHEMICAL AND HAEMATOLOGICAL PARAMETERS IN STANDARDBRED HORSES DURING A SELECTED FIELD EXERCISE TEST

Cilj ove studije je bio da se utvrde promene u koncentraciji malonil-dialdehida (MDA) u krvnoj plazmi kao i hematoloski i biohemijski profil kod 10 klinicki zdravih americkih kasaca podvrgnutih odabranim testovima opterecenja na terenu. U krvnom serumu je određivan i odnos između koncentracije MDA, krajnjeg produkta glavne lipidne peroksidacije i aktivnosti enzima misica, kreatin kinaze (CK) i aspartat aminotransferaze (AST). Uzorski venske krvi za analize su prikupljani u mirovanju, neposredno po zavrsetku opterecenja a zatim i 24 i 48 sati nakon toga. Neposredno po zavrsetku opterecenja, utvrđene su znacajne promene vecine biohemijskih i hematoloskih parametara i one se mogu smatrati normalnim fizioloskim odgovorom. Vecina ovih parametara se vratila na vrednosti registrovane u mirovanju u roku od 48 sati. Koncentracija MDA u krvnoj plazmi je odmah posle napora bila povecana, ali ovo povecanje nije bilo statisticki znacajno za razliku od vrednosti registrovanih 24 sata posle vežbe. Maksimalna vrednost MDA je registrovana nakon 48 sati. Ovaj oksidativni stres je bio indukovan opterecenjem sto potvrđuje povecan stepen lipidne peroksidacije. Na osnovu brzog pada aktivnosti CK u serumu po zavrsetku opterecenja i odsustvu znacajne korelacije između koncentracije MDA i aktivnosti misicnih enzima u serumu, može se zakljuciti da odabrani testovi opterecenja nisu imali za posledicu trajne alteracije u misicnim celijama i da nisu dovodili do njihovog ostecenja. AB The aim of the present study was to determine changes in plasma malondialdehyde (MDA) concentration and haematological and biochemical profiles in 10 clinically healthy standardbred horses subjected to a selected field exercise test. Correlations between plasma MDA, the main lipid peroxidation end-product, and muscle enzymes: creatine kinase (CK) and aspartate aminotransferase (AST) were determined in serum samples. Venous blood samples for determination of selected blood parameters were collected, immediately post exercise, and 24 and 48 hours post exercise. Significant changes in most of the biochemical and haematological parameters determined immediately after exercise reflect the normal physiological response of horses to a selected field exercise test. Most of these parameters returned to or close to the stall values within 48 hours. The concentration of plasma MDA increased immediately post exercise, though not significantly; however it increased significantly 24 hours post exercise and reached its highest value 48 hours post exercise. Thus exercise-induced oxidative stress is evidenced by increased lipid peroxidation. From the rapid decline of serum CK activity post exercise and the absence of significant correlations between MDA and serum muscle enzymes, we concluded that the selected field exercise test caused no permanent alteration in muscle cell integrity or muscle damage.

Superoxide Dismutase: A Predicting Factor for Boar Semen Characteristics for Short-Term Preservation

Superoxide dismutase (SOD), total antioxidant capacity (TAC), and thiobarbituric acid reactive substances (TBARS) in seminal plasma were evaluated on the basis of receiver operating characteristics (ROC) analysis as predictors for distinguishing satisfactory from unsatisfactory boar semen samples after storage. SOD on day 0 correlated significantly with progressive motility (r = −0.686; P < 0.05) and viability (r = −0.513; P < 0.05) after storage; TBARS correlated only with motility (r = −0.480; P < 0.05). Semen samples that, after 3 days of storage, fulfilled all criteria for semen characteristics (viability > 85%, motility > 70%, progressive motility > 25%, and normal morphology > 50%) had significantly lower SOD levels on the day 0 than those with at least one criterion not fulfilled (P < 0.05) following storage. SOD levels of less than 1.05 U/mL predicted with 87.5% accuracy that fresh semen will suit the requirements for satisfactory semen characteristics after storage, while semen with SOD levels higher than 1.05 U/mL will not fulfill with 100% accuracy at least one semen characteristic after storage. These results support the proposal that SOD in fresh boar semen can be used as a predictor of semen quality after storage.

Prevalence of antibodies against Leptospira sp. in snakes, lizards and turtles in Slovenia.

BackgroundLeptospiral infections in poikilothermic (cold blooded) animals have received very little attention and the literature concerning natural infections of these animals is limited. The aim of this study was to determine the prevalence of leptospiral antibodies in reptiles, imported into Slovenia and intended to be pets in close contact with humans. A total of 297 reptiles (22 snakes, 210 lizards and 65 turtles) were tested for specific antibodies against serovars of Leptospira interrogans sensu stricto using the microscopic agglutination test (MAT). Live cultures of different serovars were used as antigens. MAT was performed according to standard procedures and the degree of reaction was interpreted by estimating the percentage of agglutinated leptospires. Samples showing titres of ≥ 50 against one or more serovars were considered as positive.ResultsAntibodies against seven pathogenic serovars of L. interrogans sensu stricto were detected in 46 of 297 reptiles. Among 22 snakes, specific antibodies against pathogenic serovars of three Leptospira species (L. interrogans, L. kirschneri and L. borgpetersenii) at titre levels from 1:50 to 1:400 were detected in 6 snakes. In 31 of 210 lizards, specific antibodies were found in titres from 1:50 to 1:1000 and, finally, among 65 turtles (terrapins and tortoises), 9 had specific antibodies at titre levels between 1:50 and 1:1600. Animals imported from non-EU countries showed significantly higher prevalence (25.0%; 95 confidence interval: 16.7–33.3%) than animals from EU member states (10.4%; confidence interval: 6.1–14.7%).ConclusionsReptiles may be considered as potential reservoirs of L. interrogans sensu stricto. Origin of the animals is a risk factor for presence of leptospiral antibodies, especially in lizards. Special attention should be focused on animals from non-EU member states.

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.

VALUE OF SEMEN PARAMETERS, WITH SPECIAL REFERENCE TO TNF-α, IN PREDICTING THE QUALITY OF BOAR SEMEN AFTER SHORT-TERM STORAGE

The aim of this study was to address the question whether changes in boar semen quality after short-term storage could be predicted on the basis of standard semen parameters and TNF-α level determined on the day of semen collection under commercial conditions. Progressive motility showed the highest positive correlation with morphology on day 0 of collection, and progressive motility on day 3 (P < 0.05) showed a negative correlation with acrosome abnormalities (P < 0.05). According to the area under receiver operating characteristics (ROC) curves (AUCs), progressive motility could also be used in predicting semen quality after 3 days of storage (AUC > 0.5; P < 0.05). TNF-α in seminal plasma is the only parameter measured on day 0 to show a significant correlation with the percentage of viable spermatozoa after 3 days of semen storage (r = 0.495, P < 0.05). ROC analysis shows that TNF-α level is helpful in discriminating viability outcome after semen storage (AUC = 0.94, P < 0.001). We can predict with 92.35% certainty that fresh semen samples with more than 150 pg/ml of TNF-α in the seminal plasma will retain more than 85% of viable spermatozoa after 3 days of storage. Thus, TNF-α can contribute to predicting the quality of short-term stored semen.

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.

Animal and human dentin microstructure and elemental composition

Animal teeth are a common model in studies on dentin adhesive materials. The aim of this study was to compare microstructural parameters (density and diameter of dentinal tubules (DT), peritubular dentin (PTD) thickness, PTD and intertubular dentin (ITD) surface area) and chemical characteristics of canine, porcine, equine, and human root dentin. The middle layers of dentin were harvested just below a cemento-enamel junction from incisors and investigated by means of scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDXS). SEM evaluation of the specimens revealed, that porcine dentin shared most similarities with human dentin. When comparing the density of DTs, canine dentin was also found to be similar to human dentin. Elemental composition of the root dentin did not differ significantly in porcine, equine and human dentin, but in canine dentin higher magnesium value in PTD compared to ITD was found. It is known that microstructural and chemical characteristics affect the strength of the adhesive bonds created among restorative materials and dentin. According to the results of this study, porcine dentin seems to be the most appropriate model to study dental materials to be used in human restorative dentistry.