Thymus satureioides and Origanum majorana essential oils improve the quality of Beni Arouss buck semen during storage at 4°C.

This study aims to investigate the effects of essential oils (EOs), extracted from Thymus satureioides (TS) and Origanum majorana (OM), on Beni Arouss buck semen quality stored in skimmed milk at 4°C. EOs were extracted by hydro-distillation, and the chemical compounds were determined. Ejaculates were collected from six Beni Arouss bucks, once a week for 10 weeks, and they were pooled, divided into five equal aliquots and diluted to 400 × 106 sperm/ml with skimmed milk supplemented with 0.01% of OM EO, 0.01% of TS EO, 0.05% of OM EO and 0.05% of TS EO. Non-supplemented skimmed milk was considered as a control. Semen motility, kinematic parameters, viability, abnormality, membrane integrity and lipid peroxidation were evaluated at 0, 4, 8, 24, 28, 32 and 48 hr of liquid storage at 4°C. The main EO components were carvacrol (31.7%), thymol (28.0%) and borneol (14.4%) for TS, and terpinene-4-ol (31.2%), γ-terpinene (17.4%) and α-terpinene (12.7%) for OM. The results highlighted a dose-dependent effect of TS and OM EOs on all semen quality parameters. 0.01% of both EOs had a beneficial effect on the sperm preservation stored at 4°C compared with control (p < .05) excepted for the straight-line velocity. The 0.05% EO addition had harmful effects during storage particularly for TS EO. In conclusion, 0.01% of TS and OM EOs are recommended to improve the Beni Arouss buck semen preservation at 4°C.

Prevalence of naturally occurring cartilage defects in the ovine knee.

OBJECTIVE: To determine the prevalence, anatomical location and severity of cartilage defects in the stifle (knee) within a population of adult ewes (N = 65). MATERIALS AND METHODS: Articular cartilage (AC) of the distal femur, proximal tibia and patella was assessed using Osteoarthritis Research Society International (OARSI) recommendations for macroscopic and microscopic scoring of ovine cartilage. Synovial fluid analysis and histology of the synovial membrane were performed. All limbs were examined by computed tomography. RESULTS: Twenty-eight sheep (n = 28; 43%) presented at least one score 2 or score 3 lesion. Twenty-two (n = 22; 34%) sheep were macroscopically normal. Most frequent localizations of lesions were: axial aspect of the central third of the medial tibial condyle (32.7% of the lesions), middle third of the medial femoral condyle (29.4%), middle third of the articular surface of the patella (9.8%), and axial aspect of the central third of the lateral tibial condyle (9.8%). Grade of macroscopic lesions was significantly (H (3) = 29.31, P 0.000) affected by age. Macroscopic score correlated well with histological changes that can be found in osteoarthritis (OA) (r 0.83; P 0.000). Neither clinical signs of OA, nor cytological and histological signs of inflammation were identified, while imaging abnormalities were very rare. CONCLUSIONS: Our data seem to indicate that naturally occurring OA exists in ageing sheep, at least subclinically. It might be useful to take into account prevalent cartilage defects at baseline in studies using ovine models.

Upper airway resistance: species-related differences.

In veterinary medicine, upper airway resistance deserves a particular attention in equines athletes and brachycephalic dogs. Due to the anatomical peculiarities of the upper airway and/or pathological conditions, significant alterations of performance and/or well being might occur in horses and dogs. Physiological specificities and pathological changes of the lower respiratory tract deserve a major attention in other species.

Relationship between leptin content, metabolic hormones and fat deposition in three beef cattle breeds.

The aim of the study was to determine if cattle breeds differing in their carcass characteristics also differ in the profiles of their leptin and metabolic hormones. Three breeds, Belgian Blue (BB) (n=12), Limousin (L) (n=12) and Aberdeen Angus (AA) (n=12) with varying ability to deposit fat and protein were compared. Blood, muscle and subcutaneous (SC) adipose tissue were sampled. Animal performance, carcass and meat characteristics were determined as well as plasma leptin concentration, leptin gene expression in SC adipose tissue, leptin-receptor gene expression in SC adipose tissue and plasma concentration of insulin, tri-iodothyronin (T3), thyroxin (T4) and cortisol. The BB bulls showed the lowest values of leptin gene expression (P<0.05). Values of plasma leptin concentration and of leptin-receptor gene expression tended to be lower in BB than in the other breeds. For a similar amount of adipose tissue (after normalisation), BB bulls showed a higher ratio of plasma leptin (P<0.05), whereas normalised leptin gene and leptin-receptor gene expressions did not significantly differ between breeds. Belgian Blue bulls also differed in their metabolic hormone profile, tending to show lower values of insulin, T3 and T4 than the two other breeds. Cortisol levels were significantly lower (P<0.05) in BB than in L and AA animals.

Pregnancy-associated glycoprotein secretion in North Moroccan goats.

The plasma profile of pregnancy-associated glycoprotein (PAG) and its relationship with fetal number were studied in 14 North Moroccan goats using a semi-heterologous radioimmunoassay (RIA). Peripheral blood was collected every day during the first month of pregnancy, afterward the blood samples were collected trice a week. The PAG were first detected at day 20 of pregnancy, their levels increase to week 8 of gestation then decrease slightly until parturition. Statistical differences between goats carrying one or two foetuses are observed from week 5 of pregnancy until parturition. Factorial Discriminant Analysis provides mathematical models for the discrimination between the litter size using the PAG level and the week of pregnancy. Using only one blood sample per week, high sensitivity, specificity and overall accuracy (74%, 88% and 81%) were obtained using these models. It is concluded that the PAG RIA is an effective tool for early diagnosis of pregnancy and for discrimination between the litter sizes in North Moroccan goats.

Exploratory investigation of Q fever in apparently healthy meat sheep flocks in Belgium.

Q fever is a cosmopolitan disease affecting both humans and many animal species. Although sheep are often implicated in human Q fever outbreaks, the disease remains largely underestimated in meat sheep flocks. In order to fulfil this gap, a preliminary study was performed aiming to investigate the serological and molecular aspects of infection with Coxiella burnetii among meat sheep flocks in Belgium. Five Belgian sheep flocks were recruited for this work. Indirect ELISA was used, and in addition, real-time PCR was performed on samples of milk, rectal and vaginal swabs, to understand the dynamics of bacterial shedding. Despite the low overall apparent seroprevalence of 1.39% (95% CI: 0.04-7.5), a high rate of bacterial shedding was found, with 27.7% of tested sheep (N = 72) with a positive result to PCR, especially through the rectal and vaginal routes and in seronegative animals. Furthermore, Coxiella burnetii DNA was detected in 26.76% of seronegative animals. It can be concluded that an overall good clinical condition of the sheep cannot be used to exclude the presence of C. burnetii in a flock. Furthermore in the diagnosis of Q fever in sheep, serology alone was not a sensitive diagnostic tool. On the contrary, molecular biology allowed to detect bacterial shedding, which is an essential element in order to assess the risk due to the contact with shedding animals. At the light of these results, the role of meat sheep flocks in the epidemiology of Q fever in Belgium needs to be better understood.

Effects of exercise and oral antioxidant supplementation enriched in (n-3) fatty acids on blood oxidant markers and erythrocyte membrane fluidity in horses.

The aim of this study was to investigate in a placebo-controlled field study the effect of a (n-3)-vitamin supplementation on erythrocyte membrane fluidity (EMF), oxidant/antioxidant markers and plasmatic omega3/omega6 fatty acid ratio (FAR) in 12 eventing horses. Venous blood was sampled at rest before (PRE) and after (POST) a three week treatment period with either the supplement (group S, n=6) or a placebo (group P, n=6) as well as after 15min (POST E15') and 24h (POST E24h) after a standardised exercise test. The following markers were analysed: EMF, plasma antioxidant capacity of water and lipid soluble components, ascorbic acid, uric acid (UA), glutathione (reduced: GSH, oxidised: GSSG), vitamin E (Vit E), beta-carotene, glutathione peroxidase (GPx) and superoxide dismutase (SOD) activity, selenium, copper (Cu), zinc (Zn), oxidised proteins (Protox), lipid peroxides (Pool) and FAR. EMF did not differ between group S and P after treatment, but GPx remained unchanged in group S whereas it decreased in group P and plasma Cu/Zn ratio remained unchanged whereas it increased in group P. FAR were significantly increased in group S. Exercise induced a significant decrease of EMF (POST vs. E24h) in both groups, but which was significantly lower at E15' in group S than in group P. Exercise induced a significant increase of UA and ACW (POST vs. E15') and Protox (POST vs. E24h) in both groups. An exercise-related decrease in GSH and Pool (POST vs. E15') was found in group P, whereas Vit E and FAR (POST vs. E24h) significantly decreased in both groups. The study showed that exercise induced a decrease in EMF in horses associated with changes of blood oxidative balance. The (omega-3)-vitamin supplementation tested improved the oxidative balance poorly but delayed the exercise-induced decrease of EMF and increased the FAR.

Angiographic Anatomy of External Iliac Arteries in the Sheep.

External iliac artery atherosclerotic disease and aneurism occur in man. For treatment, imaging is required to facilitate minimally invasive introduction and advancement of stents within the intended vessels. Sheep are commonly used to test and improve stents. However, little information is published regarding the angiographic anatomy of the iliac arteries in the ovine species. The objective of this study was to describe the angiographic anatomy of the iliac arteries in the sheep. Computed tomography (CT) angiography and gross anatomical dissection were performed in, respectively, 10 and 43 adult ewes. Diameters and lengths of the arteries were measured. In comparison with man, salient anatomical differences were identified in the sheep: (1) the absence of common iliac arteries, (2) the common trunk at the origin of internal iliac arteries and (3) the location of the bifurcation of the external iliac arteries into femoral arteries in the pelvis (not in the limb). External iliac arteries in this series of sheep were 86 mm long in average and had a mean diameter of 7.5 mm. Lengths of arteries are only slightly different between man and sheep, while diameters are rather similar. Therefore, the sheep model appears to be sufficiently similar to man to test stent properties. This study provides useful reference images and measures of lengths and diameters of relevant arteries that could be applied to research with ovine models.

Effect of oral antioxidant supplementation on blood antioxidant status in trained thoroughbred horses.

The oxidant/antioxidant equilibrium of trained thoroughbred horses (n = 40) was assessed on three occasions during a period of three months under field conditions by blood antioxidant markers analysis, i.e. plasma ascorbic acid (AA), plasma antioxidant capacity of water-soluble components (ACW), whole blood (GSH) and oxidised (GSSG) glutathione, plasma alpha-tocopherol and beta-carotene, plasma antioxidant capacity of lipid-soluble components (ACL), red blood cell superoxide dismutase (SOD) and glutathione-peroxidase activity (GPx) and plasma trace-elements, i.e. selenium (Se), copper (Cu), zinc (Zn). A control group of ten horses receiving a placebo and an antioxidant group of 30 horses orally supplemented with an antioxidant mixture were randomly formed. An antioxidant imbalance was observed after three months in the control group, reflected by a significant decrease in GSH, SOD, GPx, Se (P < 0.05) and a significant increase in GSSG (P < 0.05). The antioxidant supplement prevented GPx and Se decrease and significantly increased ACW, alpha-tocopherol, beta-carotene and ACL (P < 0.05). Significant sex- or age-related differences were found for AA, ACW, alpha-tocopherol, SOD, GPx and Se, and there were significant correlations between ACW-AA, ACL-alpha-tocopherol, GPx-Se, CPK-Se, CPK-alpha-tocopherol and CPK-Cu. This field study has shown that trained thoroughbred horses undergo significant changes of several blood antioxidant markers and that oral antioxidant supplementation might partially counterbalance these changes by improving the hydrophilic, lipophilic and enzymatic antioxidant blood capacity.

Effect of Moxidectin Treatment at Peripartum on Gastrointestinal Parasite Infections in Ewes Raised under Tropical Andes High Altitude Conditions.

This study tested the impact of moxidectin at peripartum on nematode fecal egg count (FEC) and clinical parameters on ewes in the high altitude tropical Andes of Colombia. FEC and clinical evaluations were performed on 9 occasions in 43 naturally infected ewes before and during gestation and after lambing. Moxidectin (Mox, 200 µg kg(-1)) was applied at late pregnancy (T 1, n = 15) or 48 hours after parturition (T 2, n = 14). 14 untreated ewes served as controls (C). Suckling lambs (n = 58) remained untreated and underwent four clinical and parasitological evaluations until 8 weeks after birth. Mox efficacy equaled 99.3% (T 1) and 96.9% (T 2). Highest mean FEC value reflecting periparturient nematode egg rise (PPER) was recorded in C ewes at 4-6 weeks after lambing. Significant FEC reductions were found in T 1 (94.8%) and T 2 (96.7%) ewes (p < 0.05). All lambs showed a significant and ewes-group independent increase in FEC before weaning (p < 0.05). Clinical parameters (anemia and diarrhea) showed time- and treatment-related differences (p < 0.05). Monitoring of FEC and clinical parameters linked to gastrointestinal parasite infections allowed demonstrating that postpartum or preweaning are two critical periods to nematode infection for sheep raised under tropical Andes high altitude conditions. Use of Mox as anthelmintic treatment prevented PPER.

Impaired accumulation of granulocytes in the lung during ozone adaptation.

Respiratory alterations induced by an acute exposure to ozone (O(3)) paradoxically resolve during multiday exposure. This adaptation is characteristically accompanied by a gradual attenuation of lung neutrophilia. As maintenance of neutrophilia at the site of inflammation is due to cytokine-mediated delayed neutrophil apoptosis, which is associated with reduced levels of Bax, a proapoptotic protein, we sought to determine whether defects in these mechanisms could account for O(3) adaptation. Lung granulocytes obtained at different time points from calves exposed to 0.75 ppm O(3) for 12 h/d for 7 consecutive days neither showed enhancement of survival nor Bax deficiency, when compared to blood granulocytes. To further investigate the effects of an exogenous oxidative stress on neutrophil survival, human granulocytes were treated with hydrogen peroxide alone, or in combination with granulocyte/macrophage colony-stimulating factor, an antiapoptotic cytokine. Both treatments led to rapid apoptosis associated with downregulation of Bcl-x(L) and Bcl-2, two antiapoptotic proteins. This study shows that O(3) adaptation is associated with a failure in the mechanisms leading to accumulation of neutrophils at the site of inflammation, and suggests that this defect is due to direct proapoptotic effects of exogenous oxidative stress on granulocytes.

p65 Homodimer activity in distal airway cells determines lung dysfunction in equine heaves.

Nuclear factor-kappaB (NF-kappaB) activity, which is a key regulator of inflammatory gene expression, is increased in bronchial epithelial cells from horses suffering from heaves (a hypersensitivity-associated inflammatory condition of the lung). To determine whether this increased activity extends to distal airways and to other pulmonary cells, cells recovered by broncho-alveolar lavage (BAL) in healthy and heaves-affected horses were assessed for NF-kappaB activity. NF-kappaB activity was much higher in BAL cells from heaves-affected horses, especially during crisis (disease exacerbation), than in cells from healthy horses. Moreover, the level of NF-kappaB activity found in BAL cells was positively correlated to total lung resistance and to the proportion of neutrophils present in BAL fluid. Finally, prototypical p65-p50 NF-kappaB heterodimers were absent from BAL cells, which mostly contained p65 homodimers. These results (1) show that increased NF-kappaB activity is a general feature of heaves lung; (2) demonstrate the importance of p65 homodimers in neutrophilic inflammation; and (3) suggest that the use of specific NF-kappaB inhibitors could improve lung function in heaves-affected horses.

Indices of oxidative stress in blood and pulmonary epithelium lining fluid in horses suffering from recurrent airway obstruction.

To test the hypothesis that reactive oxygen species could be associated to the lower airway disorders occurring in horses suffering from recurrent airway obstruction (RAO), indices of oxidative stress were studied in blood and pulmonary epithelium lining fluid in 5 RAO horses either in clinical remission or 24 h after the onset of a crisis of bronchospasm and in 5 healthy horses. Venous blood and bronchoalveolar lavage fluid (BALF) samples were collected and analysed for reduced glutathione (GSH), oxidised glutathione (GSSG), total glutathione (TGSH), glutathione redox ratio (GRR) in blood haemolysate and pulmonary epithelium lining fluid (PELF). The haemolysate concentrations of GSH, GSSG, TGSH and GRR were similar in the 3 groups. The PELF glutathione status was significantly different in the RAO horses in acute crisis compared to healthy horses, indicating the occurrence of an oxidative stress. When RAO horses were in crisis their GSH and TGSH remained unchanged but their GSSG and GRR were significantly increased compared to the remission. These results support the hypothesis that oxidative stress is associated with lower airway disorders occurring in horses suffering from RAO.

The use of cardboard bedding material as part of an environmental control regime for heaves-affected horses: in vitro assessment of airborne dust and aeroallergen concentration and in vivo effects on lung function.

This study aimed to test whether shredded cardboard is an appropriate minimum-dust bedding material for heaves-affected horses. Results of standardized in vitro measurement of airborne dust and aeroallergen concentrations of cardboard bedding were significantly lower than those of common bedding materials. Six heaves-affected horses in clinical remission after pasturing were stabled for two months on cardboard bedding and fed grass silage. Pulmonary function tests (PFT: ventilatory mechanics, arterial blood gases, airway inflammation scoring, bronchoalveolar cytology) were performed before, during and after this period and after stabling the horses in poor hygienic conditions. PFT values measured during and after the stabling period on cardboard bedding were not significantly different from those recorded after the period at pasture or from those of healthy horses, but were significantly different from those recorded in poor hygienic conditions. On basis of the in vitro and in vivo results it can be concluded that cardboard bedding, used in conjunction with low-dust forage, may be appropriate in the provision of minimum-dust management of heaves-affected horses.

Influence of breathing pattern and lung inflation on impulse oscillometry measurements in horses.

The objective of this paper was to determine if changes in ventilation patterns could influence the outcome of respiratory function measurements performed with our impulse oscillometry system (IOS) in horses. In a first study, IOS tests were performed in vitro on six isolated equine lungs. Lung inflation levels were controlled by modifying depressurisation inside an artificial thorax and different ventilation patterns were imposed. In a second in vivo study, transient variations in breathing pattern were evaluated both with the IOS and a current reference technique (CRT) in five healthy mature horses after an intravenous (i.v.) injection of lobeline hydrochloride. In both studies, respiratory rate (RR, range: 7-42 breaths/min.) and tidal volume (V(T), range: 0.4-25 L) had minor or no influence on IOS parameters. The influence of lung inflation, most marked for resistance at 5 Hz (R(5 Hz)), was limited for the considered physiological range. In vivo, statistical models indicated that maximal changes in pleural pressure (Max Delta Ppl) and peak flows were the main determinants of the variability of the resistance (R(rs)) and the reactance (X(rs)) of the respiratory system. The fourfold increase in baseline Max Delta Ppl and peak flows obtained during hyperpnoea caused a significant increase in R(rs) at 5 and 10 Hz and a decrease in X(rs) at all frequencies. We conclude that IOS parameters are not influenced by tachypnoea, but will reflect alterations in respiratory mechanics caused by hyperpnoeic breathing.

Effect of chronic airway inflammation and exercise on pulmonary and systemic antioxidant status of healthy and heaves-affected horses.

In heaves-affected horses the relation between oxidant status, airway inflammation (AI) and pulmonary function (PF) is unknown. The oxidant status of blood and pulmonary epithelial lining fluid (PELF) of healthy (H, n = 6) and heaves-affected horses in clinical remission (REM, n = 6) and in crisis (CR, n = 7) was assessed at rest, during and after standardised exercise test by measurement of reduced and oxidised glutathione, glutathione redox ratio [GRR%]; uric acid and 8-epi-PGF2alpha. Oxidant status was related to PF parameters (mechanics of breathing and arterial blood gas tension) and Al parameters (bronchoalveolar lavage [BAL] neutrophil % and AI score). Haemolysate glutathione was significantly different between groups and was correlated with PF and AI parameters; GRR in PELF was increased during CR and was correlated with PF and AI parameters. Exercise induced an increase of plasma uric acid that was significantly higher both in REM and CR. PELF 8-epi-PGF2alpha was significantly increased in CR and correlated with PF and AI parameters. These results suggest that oxidative stress occurring in heaves is correlated with PF and AI and may be locally assessed by PELF glutathione status, uric acid and 8-epi-PGF2alpha. Systemic repercussions are reflected by assay of GSH in resting horses and by uric acid in exercising horses.

Effect of nutritional antioxidant supplementation on systemic and pulmonary antioxidant status, airway inflammation and lung function in heaves-affected horses.

An oxidant/antioxidant imbalance in favour of oxidants has been identified as playing a decisive role in the pathogenesis of chronic inflammatory airway diseases. Nutritional antioxidant supplementation might reduce oxidative damage by enhancement of the antioxidant defence, thereby modulating inflammatory processes. In a placebo-controlled, blind study, it was tested whether a dietary antioxidant supplement administered for 4 weeks would improve lung function and reduce airway inflammation in heaves-affected horses. Eight horses in clinical remission of heaves were investigated at rest and after a standardised exercise test before and after treatment with an antioxidant supplement (consisting of a mixture of natural antioxidants including vitamins E and C and selenium from a variety of sources) or placebo (oatfeed pellets without additive). Pulmonary function and exercise tolerance were monitored; systemic and pulmonary lining fluid uric acid, glutathione and 8-epi-PGF(2alpha) were analysed, and bronchoalveolar lavage (BAL) cytology and inflammatory scoring of the airways were performed. The antioxidant treatment significantly improved exercise tolerance and significantly reduced endoscopic inflammatory score. Plasma uric acid concentrations were significantly reduced, suggesting downregulation of the xanthine-dehydrogenase and xanthine-oxydase pathway. Haemolysate glutathione showed a nonsignificant trend to increase, while plasma 8-epi-PGF(2alpha) remained unchanged. Pulmonary markers and BAL cytology were not significantly affected by antioxidant supplementation. The present study suggests that the antioxidant supplement tested modulated oxidant/antioxidant balance and airway inflammation of heaves-affected horses.

Preliminary survey on the impact of Schmallenberg virus on sheep flocks in South of Belgium.

Between late February and May 2012, a preliminary anonym survey was conducted among sheep farmers in south of Belgium in order to contribute to future estimations of the economic losses caused by Schmallenberg virus (SBV). Based on clinical signs consistent with SBV infection, this survey involved 13 meat sheep flocks considered as positive flocks with subsequent SBV detection by RT-qPCR [SBV-positive flocks (PF); total of 961 animals], and 13 meat sheep flocks considered as negative flocks (NF; total of 331 animals). These preliminary results indicated several significant characteristics that were more present in PF than in NF. These include an increased rate of abortions (6.7% in PF versus 3.2% in NF), of lambs born at term but presenting malformations (10.1% in PF versus 2.0% in NF) and of dystocia (10.1% in PF versus 3.4% in NF). Lamb mortality during the first week of life was reported more frequently in PF (8 of 13 PF, 61.5%) than in NF (1 of 13 NF, 7.7%). In PF, the observed prolificacy rate was 2-fold lower (93%) than expected (186%). The implementation of a survey at larger scale, including a high number of breeders, is necessary to allow a more detailed analysis of the SBV impact in the sheep sector.

Field veterinary survey on clinical and economic impact of Schmallenberg virus in Belgium.

We implemented a questionnaire-based methodology targeting veterinary field practitioners to evaluate clinical and economic impact of Schmallenberg virus in Belgium. First suspicious cases were detected as soon as July 2011. The mean cost for individual symptomatic treatment was 65 or 107 Euros, in case of fatal outcome or apparent recovery, respectively.