Annick Gabriel

Comparative pathogenesis of acute and latent infections of calves with bovine herpesvirus types 1 and 5

Summary. This study was conducted to compare the pathogenesis of acute and latent infections with closely related bovine herpesvirus types 1 (BHV-1) and 5 (BHV-5) in their natural host. Two groups of eight calves were inoculated intranasally with BHV-1 or BHV-5. Although BHV-1 and BHV-5 similarly replicate in the nasal mucosa after inoculation, both viruses differ markedly in their ability to cause disease, BHV-5 being responsible of some fatal encephalitis while BHV-1 inducing rhinotracheitis. Virus isolation and immunohistochemistry demonstrated that BHV-5 replicates extensively in neurons of the central nervous system (CNS) and in respiratory cells of lungs, tracheal and nasal mucosae. Invasion of the CNS likely occurs through the trigeminal and olfactory pathways. Both groups developed cross-neutralising antibodies during this experiment suggesting partial clinical cross-protection afforded by the two infections. Three months after primary infection, experimental reactivation showed that BHV-5 was able to establish latency in the trigeminal ganglia but also the CNS of surviving calves. Moreover, laboratory findings suggested that BHV-5 could also persist in the tracheal and nasal mucosae. These results indicate that, after primary infection, BHV-1 and BHV-5 displayed similar biological features and consequently need to be considered together for the control of BHV-1 infection.

Latency and reactivation of a glycoprotein E negative bovine herpesvirus type 1 vaccine: influence of virus load and effect of specific maternal antibodies.

The effects of the vaccination of neonatal calves with a glycoprotein E (gE)-negative bovine herpesvirus type 1 (BHV-1) were investigated in naïve and passively immunised calves either with the recommended dose or a 5-fold concentrated one. After inoculation (PI), all calves excreted the virus vaccine except three passively immunised calves inoculated with the lower titre. No antibody response could be detected in passively immunised calves, whatever the dose used, and they all became BHV-1 seronegative and remained so after dexamethasone treatment (PDT). Nevertheless, as shown by a gamma-interferon assay, all calves that excreted the vaccine PI developed a cell-mediated immune response and a booster response was observed PDT, suggesting viral reactivation. The vaccine virus was recovered PDT from nasal secretions in two calves and BHV-1 DNA were detected in trigeminal ganglia from five calves belonging to all inoculated groups. The results show that the BHV-1 gE-negative vaccine can establish latency not only in naïve but also in passively immunised neonatal calves after a single intranasal inoculation. Moreover, this study shows for the first time that the gE-negative vaccine, when used in passively immunised calves, can lead to seronegative vaccine virus carriers.

The relationship between carcass characteristics, plasma hormones and metabolites in young fattening bulls.

Six Belgian Blue bulls (double-muscled type) and six Friesian bulls were offered a fattening diet for 34 weeks. Plasma samples were obtained once a week and also every 20 min over a 24 h period, 7 weeks before slaughter.No differences were observed between the breeds in plasma glucose, urea and free amino nitrogen concentrations, while creatinine was significantly higher in the Belgian Blue bulls. Tri-iodothyronin, tetra-iodothyronin, insulin-like growth factor 1, insulin and testosterone concentrations were higher in the Holstein group. In contrast, the Belgian Blue bulls appeared to produce more growth hormone. The slaughter weight, carcass weight, dressing percentage and proportion of lean meat were significantly higher in the Belgian Blue group. The characteristics of muscle mass (carcass weight, dressing percentage and proportion of lean meat) were positively correlated with creatinine and with the total peak area or peak amplitude of growth hormone. The insulin concentration was positively correlated with the proportion of adipose tissue in the carcass and negatively correlated with the proportion of muscle. There were no correlations between the carcass characteristics and insulin-like growth factor 1 or testosterone. No further information was provided when the ratios of the hormones were correlated with carcass characteristics.

Effects of dietary carbohydrate composition on rumen fermentation, plasma hormones and metabolites in growing-fattening bulls.

Two groups of six bulls were offered a fattening diet based on barley or sugar-beet pulp in the proportions of either 500: 200 g/kg or 200: 500 g/kg to compare the effects of starch and degradable fibre on rumen fermentations, plasma hormones and metabolites. The bulls were fed twice daily at 08.00 and 16.00 h. The large proportion of sugar-beet pulp in the diet induced higher levels of acetic and butyric acids and less propionic acid than the high proportion of barley. The diurnal patterns over a 24-h period of rumen pH, volatile fatty acids, glucose and ammonia were characterized by two cycles. Sugar-beet pulp in large proportions induced more even fermentations as indicated by flatter curves with less extreme values than those produced by a large inclusion of barley in the diet. Ammonia concentration was high before feeding and decreased subsequently during 4 to 6 h after the meal. No typical patterns were observed in plasma concentrations of glucose and alpha-amino nitrogen. Plasma urea concentration rose 2h after feeding and dropped during the following 6 to 8 h. There were no effects of dietary carbohydrate composition on growth hormone concentration but plasma insulin was significantly reduced with the diet high in sugar-beet pulp. Furthermore, the insulin profile showed peaks after each meal with the barley diet but only after the afternoon feeding with the diet high in sugar-beet pulp.

FEASIBILITY OF ULTRASOUND-GUIDED EPIDURAL ACCESS AT THE LUMBO-SACRAL SPACE IN DOGS

Epidural injections are commonly performed blindly in veterinary medicine. The aims of this study were to describe the lumbosacral ultrasonographic anatomy and to assess the feasibility of an ultrasound-guided epidural injection technique in dogs. A cross sectional anatomic atlas of the lumbosacral region and ex vivo ultrasound images were obtained in two cadavers to describe the ultrasound anatomy and to identify the landmarks. Sixteen normal weight canine cadavers were used to establish two variations of the technique for direct ultrasound-guided injection, using spinal needles or epidural catheters. The technique was finally performed in two normal weight cadavers, in two overweight cadavers and in five live dogs with radiographic abnormalities resulting of the lumbosacral spine. Contrast medium was injected and CT was used to assess the success of the injection. The anatomic landmarks to carry out the procedure were the seventh lumbar vertebra, the iliac wings, and the first sacral vertebra. The target for directing the needle was the trapezoid-shaped echogenic zone between the contiguous articular facets of the lumbosacral vertebral canal visualized in a parasagittal plane. The spinal needle or epidural catheter was inserted in a 45° craniodorsal-caudoventral direction through the subcutaneous tissue and the interarcuate ligament until reaching the epidural space. CT examination confirmed the presence of contrast medium in the epidural space in 25/25 dogs, although a variable contamination of the subarachnoid space was also noted. Findings indicated that this ultrasound-guided epidural injection technique is feasible for normal weight and overweight dogs, with and without radiographic abnormalities of the spine.

MORPHOMETRIC STUDY OF THE EQUINE NAVICULAR BONE : VARIATIONS WITH BREEDS AND TYPES OF HORSE AND INFLUENCE OF EXERCISE

Navicular bones from the 4 limbs of 95 horses, classified in 9 categories, were studied. The anatomical bases were established for the morphometry of the navicular bone and its variations according to the category of horse, after corrections were made for front or rear limb, sex, weight, size and age. In ponies, navicular bone measurements were smallest for light ponies and regularly increased with body size, but in horses, navicular bone dimensions were smallest for the athletic halfbred, intermediate for draft horse, thoroughbreds and sedentary halfbreds and largest for heavy halfbreds. The athletic halfbred thus showed reduced bone dimensions when compared with other horse types. Navicular bones from 61 horses were studied histomorphometrically. Light horses and ponies possessed larger amounts of cancellous bone and less cortical bone. Draft horses and heavy ponies showed marked thickening of cortical bone with minimum intracortical porosity, and a decrease in marrow spaces associated with more trabecular bone. Two distinct zones were observed for the flexor surface cortex: an external zone composed mainly of poorly remodelled lamellar bone, disposed in a distoproximal oblique direction, and an internal zone composed mainly of secondary bone, with a lateromedial direction for haversian canals. Flexor cortex external zone tended to be smaller for heavy ponies than for the light ponies. It was the opposite for horses, with the largest amount of external zone registered for draft horses. In athletic horses, we observed an increase in the amount of cortical bone at the expense of cancellous bone which could be the result of reduced resorption and increased formation at the corticoendosteal junction. Cancellous bone was reduced for the athletic horses but the number of trabeculae and their specific surfaces were larger. Increased bone formation and reduced resorption could also account for these differences.

Morphometric Study of the Equine Navicular Bone: Age-Related Changes and Influence of Exercise

Navicular bones from the four limbs of 95 horses, classified in 9 categories, were studied. The effects of age on navicular bone morphometry and histomorphometry were estimated, after adjustment of the data to even out the effects of front and rear limbs, morphometrical type, sex, weight, and size.All the external measurements of the navicular bone decreased significantly with increasing age. From the histomorphometrical data, cortical bone volume decreased with age in most horses, whereas cancellous bone volume and, in particular, the marrow spaces increased. The increase in the cancellous bone volume could have resulted from tunnelling of the internal part of the cortex, which converted it progressively into a porous trabecular-like structure. Trabecular bone volume also decreased with age and the trabecular lattice changed dramatically to become disconnected in aged horses. These observations corresponded closely to those reported for ageing of the skeletal system in humans.However, in sporting horses, the navicular cortical bone volume increased with age and the cancellous bone volume decreased. Exercise appeared to have decreased bone resorption and increased bone formation at the endocortical junction. The cancellous bone architecture was also improved, in that the trabecular lattice and trabecular bone volume remained unchanged in aged sporting horses. Our findings confirmed that exercise may be good practice to prevent age-related bone loss.

Morphometric analyses of the body and the branches of the normal third interosseous muscle (suspensory ligament) in Standardbreds

The third interosseous muscle (suspensory ligament, TIOM) is composed of connective tissue (CT) with a variable proportion of muscle (MT) and adipose tissue (AT). The aim of our study is to quantify the CT, MT and AT within the body and the branches of right thoracic and pelvic limbs TIOM in sound horses to determine whether there are differences in CT, MT and AT between age, sex, limbs and levels. Right limbs from 11 sound horses were collected. Samples from 6 levels of the TIOM were embedded in paraffin or in Tissue‐Tek®. Most of the paraffin sections were shredded. Using the cryosection, some artefacts appeared. Cryoprotection was carried out, which produced the best results. Hematoxylin–phloxine–saffron and Hematoxylin–eosin gave a good contrast of colours between the tissues observed allowing the use of an image analysis programme to calculate percentage of each tissue within the TIOM. The percentage of MT and AT decreased significantly (P < 0.0001), whereas the percentage of CT increased significantly (P < 0.0001) with age and when descending from the proximal to the distal level of the TIOM. The percentage of MT was significantly higher (P < 0.0001) in females than males, while the percentage of CT was significantly higher (P < 0.0001) in males than females. The percentage of AT was significantly higher (P = 0.0278) in pelvic limbs than in thoracic limbs. These results confirm the variation in tissue composition within the TIOM of sound horses.

Arterial Head Vascularization Cartographies of Normal Metencephalic Dogs Using Magnetic Resonance Angiography

The aim of our study was to establish head arterial cartographies—useful for the diagnosis of brain diseases leading to cerebral vascular modifications—by means of magnetic resonance angiography (MRA). Casts of the arterial vascular brain system were used to corroborate the MRA results as they can be easily rotated in nonvirtual three‐dimensions and give an accurate view of the arteries calibre and origin. Two types of 3T MRA images were used: three‐dimensional fast low‐angle shot (3D‐FLASH) acquisition sequenced every 20 s, paired with injection of a paramagnetic contrast medium, and three‐dimensional time‐of‐flight (3D‐TOF) acquisition sequenced every 300 s. 3D‐FLASH acquisition gives very accurate images of the cerebral arteries and veins, but must be used with care in debilitated animals. 3D‐TOF acquisition is less accurate and gives only images of the main cerebral arteries without showing the venous system. It is, however, a viable diagnostic method for monitoring vascular lesions (e.g., cerebral hemorrhages). Anat Rec, 2011.

Mechanoreceptors in the Anterior Horn of the Equine Medial Meniscus: an Immunohistochemical Approach

Lameness due to stifle and especially meniscal lesions is frequent in equine species. In humans, mechanoreceptors involved in proprioceptive function are well studied. Given the high incidence of meniscal injuries in horses, and the lack of information concerning them in equine menisci, our objective was to study these corpuscles in six healthy anterior horns of the equine medial meniscus, which is the most common localisation reported for equine meniscal injuries. Immunohistochemical stainings were performed using antibodies against high molecular weight neurofilaments and glial fibrillary acidic proteins. From a purely fundamental point of view, our work highlights for the first time the presence of Ruffini, Pacini and Golgi corpuscles in equine meniscus. They were found, isolated or in clusters and always located at the vicinity of blood vessels, at the level of the anterior horn of the equine medial meniscus. This morphological approach could serve as a basis for clinical studies, to evaluate the impact of these corpuscles on the poor sportive prognosis in equine meniscal tears.