Different action of IBMX,isoproterenol and rutin on orthovanadate-induced nitric oxide release in mouse macrophage cells

The effects of cAMP-elevating compounds IBMX (3-isobutyl-1-methylxanthine) and isoproterenol, and that of rutin (an effective superoxide scavenger) were studied on orthovanadate--(a putative protein-phosphotyrosine phosphatase inhibitor) induced nitric oxide (NO) production in J774A.1 mouse macrophage cells. As we previously reported (Koncz and Horváth, 2000), rutin and sodium orthovanadate act synergistically to induce production of high amount of NO in J774A.1 cells. IBMX, an agent that can elevate cAMP level in the cells, can reduce the production of both the LPS- and rutin + orthovanadate-induced NO in macrophages. In contrast, isoproterenol, a non-selective beta-adrenergic receptor agonist, that reduced the LPS-induced NO production in macrophage cells, was unable to reduce the rutin + orthovanadate-induced NO production without negatively affecting cell viability. Moreover, isoproterenol dramatically enhanced the orthovanadate-induced NO synthesis in J774A.1 cells. Our previous study clarified that rutin and orthovanadate, in a specific concentration ratio of both, were able to produce hydrogen peroxide (H2O2). Using 2',7'-dichlorofluorescein-diacetate as a marker for H2O2, isoproterenol alone induced its oxidation but the rutin plus orthovanadate-induced H2O2 production was reduced by isoproterenol. These observations have revealed that, in some cases, H2O2 and superoxide (O2-) scavengers can act in a reverse mode on macrophage cells depending on the presence or absence of orthovanadate.     

Influence of age and purpose for testing on the cut-off selection of serological methods in bovine neosporosis

The aim of this study was to investigate the need for different cut-off points, according to animal age and the purpose of testing, for two of the most widely used serological techniques in bovine neosporosis, IFAT and a crude antigen ELISA (Civtest, HIPRA). Therefore, the population reference sera used were defined using a combination of multiple criteria such as epidemiological/clinical and histopathological parameters and an immunoblot test. Firstly, foetuses and breeding cattle (heifers and cows) were considered as separate subpopulations for serological evaluation. Secondly, cut-off points for each serological technique (IFAT and ELISA) according to age group (foetuses and breeding cattle) and the different practical applications (detection of infection and abortion) were calculated following the receiver operating characteristics (ROC) analysis. Cut-off points were defined, for IFAT and ELISA for aborted breeding cattle and for IFAT alone in the case of the foetuses, assuming an equivalent cost of false positive and negative results. In infected breeding cattle, for IFAT and ELISA and in foetuses for ELISA, two possible cut-off values were obtained, one for a maximum sensitivity and one for a maximum specificity and the intervals of unclear results were defined. In this case, a cut-off value for equal sensitivity and specificity was also estimated. When cut-off points for infected breeding cattle, 1:100-1:250 for IFAT and 0.306-0.451 for ELISA were applied to a target population, optimal and similar negative and positive predictive values together with similar apparent and true prevalence results were observed suggesting the possibility of using both tests interchangeably.     

Reno-, hepato- and splenomegaly of common carp fingerlings (Cyprinus carpio L.) diseased in swimbladder inflammation caused by Sphaerospora renicola Dyková et Lom, 1982

The weight of internal organs (swimbladder, kidney, liver, spleen) in relation to the body weight was studied in common carp fingerlings divided into three groups on the basis of swimbladder appearance and microscopic examination of the kidney. The fish had been collected from different Hungarian fish farms at the time when swimbladder inflammation (SBI) usually occurs (in July and August). The first group comprised fish with severe signs of SBI and massive renal sphaerosporosis, the second group consisted of fish with milder swimbladder changes and/or kidney infection by a low number of Sphaerospora renicola, while the third group was constituted by infection-free common carp fry. Statistical analysis of swimbladder, kidney, liver and spleen weight in relation to the body weight revealed that in the infected groups the internal organs were substantially enlarged. This suggests that in common carp fry with SBI the swimbladder changes are accompanied by reno-, hepato- and splenomegaly.     

The surgical technique and the age of the horse both influence the outcome of mosaicplasty in a cadaver equine stifle model

Six pieces of grafts, 6.5 mm in diameter, 20 mm in length, were taken from each of 170 cadaver hindlimbs, using the cranial surface of the medial femoral trochlea for harvesting. The age of the horses varied between 4 months and 23 years. 30 limbs under the age of 12 years were selected for transplantation. Three of six grafts were transplanted into the medial femoral condyle using different combinations of tunnel depth and dilation. With ageing, a significant decline in transplantability was detected. In general, mosaicplasty cannot be recommended in horses above 11 years. Based on a previous clinical case (Bodó et al., 2000), a good surface alignment was indeed achieved with a combination of graft length drilling and dilation in most cases. However, the occasional entrapment of cartilage debris under the graft prevented perfect alignment in the present cadaver study in 27% of the grafts transplanted in this manner. Since the protrusion of grafts never exceeded 1.5 mm, we conclude that drilling 3-5 mm deeper than graft length with graft length deep dilation can avoid disadvantageous protrusion of the transplanted hyaline cartilage caps, achieving bone decompression at the same time.     

The Effect of Supplementation with Fresh Browse of Ziziphus mauritiana or Combretum aculeatum on Feed Intake,Nitrogen Utilization and Growth of Grazing Range Sheep

The influence of 0, 60 or 120 min access to a grove of either Ziziphus mauritiana (Ziziphus) or Combretum aculeatum (Combretum) on forage intake and on the digestibility, growth and excretion of nutrients was determined using 40 Oudah rams grazing for 7 h/day on poor-quality dry season pasture in the Sahel. The effects of browse species and browsing duration (0, 30, 60 or 120 min) on the ruminal ammonia content were also evaluated using 8 mature fistulated rams. Ziziphus increased both total digestible organic matter and total dry matter intakes per (kg live weight)^0.75 without decreasing herbage intake, whereas 60 or 120 min access to Combretum reduced herbage intake by 6.3% and 4.2%, respectively. The digestibility of the diet decreased (p<0.05) with the duration of access to the groves. Despite the provision of more nitrogen (N) than in the control diet, ruminal NH₃-N decreased 24 h after browsing commenced. The reduced live weight gain of sheep browsing Combretum may indicate more deleterious compounds in Combretum than in Ziziphus. Ziziphus appears to have more potential than Combretum to increase sheep production in low-input crop/livestock systems.     

Cross-sectional study on prevalence of Trypanosoma evansi infection in domestic ruminants in an endemic area of the Canary Islands (Spain)

Trypanosoma evansi is the most widely spread of the pathogenic African trypanosomes of animals. The disease (surra) was first diagnosed in the Canary Islands in a dromedary camel in 1997; thus, a control plan was implemented achieving the eventual eradication of T. evansi from most of the infected areas in the Archipelago. However, a little area remains still infected despite the use of the same control measures. To evaluate possible reservoirs in the area a representative sample of domestic ruminants was examined by serological, parasitological and molecular tests. Of a total of 1228 ruminants assessed, 61 (5%) were serologically positive (7 cattle, 21 goats, 33 sheep), but T. evansi could be demonstrated in none of them. According to FreeCalc assessment, cattle and goat populations would be free from disease; however, the results from sheep are not adequate to conclude that the population would be free from disease. As a conclusion, surveillance must be exercised on ruminant farms in the surroundings of the infected area in order to evaluate the possible extension of the disease and their potential role as reservoirs of T. evansi.     

Correlation of magnetic resonance images with anatomic features of the equine tarsus

OBJECTIVE: To correlate anatomic features of the equine tarsus identified in plastinated sections with images obtained via magnetic resonance imaging (MRI). ANIMALS: 4 horses. PROCEDURE: MRI (1.5-Tesla magnet) of the tarsus was performed on the pelvic limbs of 4 clinically normal horses following euthanasia. After imaging, tarsocrural joint spaces and vasculature were injected with colored latex. Sagittal and transverse sections of the tarsi were plastinated to facilitate interpretation of MR images. RESULTS: Relevant anatomic structures were identified and labeled on the plastinated tissue slices and corresponding MR images. Results indicated high correlations between MRI findings and those of plastinated sections. CONCLUSIONS AND CLINICAL RELEVANCE: The data obtained provided certain reference standards for normal anatomic structure sizes and positions in the equine tarsus. This information may aid future physiologic or clinical studies of this joint.     

Pharmacokinetics of danofloxacin in horses after intravenous, intramuscular and intragastric administration

REASONS FOR PERFORMING STUDY: Danofloxacin is a fluoroquinolone developed for veterinary medicine showing an excellent activity. However, danofloxacin pharmacokinetics profile have not been studied in horses previously. OBJECTIVE: To study the pharmacokinetics following i.v., i.m. and intragastric (i.g.) administration of 1.25 mg/kg bwt danofloxacin to 6 healthy horses. METHODS: A cross-over design was used in 3 phases (2 x 2 x 2), with 2 washout periods of 15 days (n = 6). Danofloxacin (18%) was administered by i.v. and i.m. routes at single doses of 1.25 mg/kg bwt. For i.g. administration an oral solution was prepared and administered via nasogastric tube. Danofloxacin concentrations were determined by HPLC assay with fluorescence detection. Tolerability at the the site of i.m. injection was monitored by creatine kinase (CK) activity. RESULTS: Danofloxacin plasma concentration vs. time data after i.v. and i.g. administration could best be described by a 2-compartment open model. The disposition of i.m. administered danofloxacin was best described by a one-compartment model. The terminal half-lives for i.v., i.m. and i.g. routes were 6.31, 5.36 and 4.74 h, respectively. Clearance value after i.v. dosing was 0.34 l/kg bwt/h. After i.m. administration, absolute bioavailability was mean +/- s.d. 88.48 +/- 11.10% and Cmax was 0.35 +/- 0.05 mg/l. After i.g. administration, absolute bioavailability was 22.36 +/- 6.84% and Cmax 0.21 +/- 0.07 mg/l. CK activity following i.m. dosing increased 3-fold over pre-injection levels 12 h after dosing and subsequently approached (but did not reach) normal values at 72 h post dose. CONCLUSIONS: Systemic danofloxacin exposure achieved in horses following i.m. administration was consistent with the predicted blood levels needed for a positive therapeutic outcome for many equine infections. Conversely, danofloxacin utility by the i.g. route was limited by low bioavailability. Tolerability associated with i.m. administration was high. POTENTIAL RELEVANCE: Pharmacokinetics, blood levels and good tolerability of i.v. and i.m. administration of danofloxacin in horses indicates that it is likely to be effective for treating sensitive bacterial infections.     

Supplementing a skimmed milk–egg yolk-based extender with L-carnitine helps maintain the motility,membrane integrity and fertilizing capacity of chilled ram sperm

This study examines the effect of L-carnitine (LC) on chilled ram semen stored for up to 96 hr. Semen samples were collected, placed in a skimmed milk + 6% egg yolk extender, pooled, aliquoted and diluted with the same extender supplemented with different LC concentration: 0 (control), 1 mM (LC1), 2.5 mM (LC2.5), 5 mM (LC5), 7.5 mM (LC7.5) or10 mM (LC10). Sperm kinetics and membranes (plasma, acrosome and mitochondrial) were examined using the CASA system and triple fluorescence staining (PI/ PNA-FITC/Mitotracker). The progressive motility was greater (p < .05) with LC7.5 treatment than the control sperm at 96 hr. The curvilinear velocity (p < .01) and the percentage of sperm with intact membranes (plasma/acrosome/mitochondria) (p < .01) were greater with all LC treatments than the control group at all times. Straight line velocity was greater (p < .01) with LC5 and LC7.5 treatments than the control group after 48 hr. The LC5 group also returned lower ALH values (p < .05) than these seen for the control groups after 48 hr. The fertilizing capacity of LC5 samples stored at 15°C for 2 hr (LC5-15°C-2h) and at 5°C for 24 hr (LC5-5°C-24h) was tested in three ewe groups via cervical fixed-time artificial insemination. In two groups, the fertilizing capacity of the LC5-5°C-24h was reduced (p < .001). In the remaining group, however, no significant difference was seen between the LC5-15°C-2h and LC5-5°C-24h sperm in this respect (pregnancy rates 52.4% versus 42.8%; p > .05). Overall, the present results suggest that supplementing skimmed milk–egg yolk-based extenders with LC has a positive effect on chilled sperm variables and fertilizing capacity.