Intervertebral disc degeneration in warmblood horses: Histological and biochemical characterization.

Gross morphology of healthy and degenerated intervertebral discs (IVDs) is largely similar in horses as in dogs and humans. For further comparison, the biochemical composition and the histological and biochemical changes with age and degeneration were analyzed in 41 warmblood horses. From 33 horses, 139 discs and 2 fetal vertebral columns were evaluated and scored histologically. From 13 horses, 73 IVDs were assessed for hydration, DNA, glycosaminoglycans, total collagen, hydroxyl-lysyl-pyridinoline, hydroxylysine, and advanced glycation end-product (AGE) content. From 7 horses, 20 discs were assessed for aggrecan, fibronectin, and collagen type 1 and 2 content. Histologically, tearing of the nucleus pulposus (NP) and cervical annulus fibrosus (AF), and total histological score (tearing and vascular proliferation of the AF, and chondroid metaplasia, chondrocyte-like cell proliferation, presence of notochordal cells, matrix staining, and tearing of the NP) correlated with gross degeneration. Notochordal cells were not seen in IVDs of horses. Age and gross degeneration were positively correlated with AGEs and a fibrotic phenotype, explaining gross degenerative changes. In contrast to dogs and humans, there was no consistent difference in glycosaminoglycan content and hydration between AF and NP, nor decrease of these variables with age or degeneration. Hydroxylysine decrease and collagen 1 and AGEs increase were most prominent in the NP, suggesting degeneration started in the AP. In caudal cervical NPs, AGE deposition was significantly increased in grossly normal IVDs and total collagen significantly increased with age, suggesting increased biomechanical stress and likelihood for spinal disease in this part of the vertebral column.

Cervical disc width index is a reliable parameter and consistent in young growing Dutch Warmblood horses.

Intervertebral disc disease, as well as the associated alteration of the radiographic intervertebral disc space width, has been reported in horses. Disc height index (DHI) has proven to be an accurate and objective parameter in other species but data related to this parameter are lacking in horses. Therefore, the aims of this retrospective longitudinal diagnostic accuracy study were (a) to evaluate the reliability of measurements within and between observers of the equine Disc Width Index (EDWI) as a parameter for radiographic equine cervical intervertebral disc space width, and (b) to evaluate the sequential development of the EDWI over time. For this, EDWI from all intervertebral disc spaces between second cervical (C) to first thoracic (Th) vertebrae were obtained in a group of 39 Dutch Warmblood horses at 1, 5, and 18 months of age, by one European College of Veterinary Diagnostic Imaging (ECVDI) board-certified veterinary radiologist (S.V.) and two veterinary students. Bland-Altmann plots and intraclass Correlation Coefficient revealed a good intra- and interobserver agreement. A linear mixed-effect model did reveal that mean EDWI increases significantly toward the caudal cervical spine, but did not differ significantly for a certain location over time or between sexes. Spearman's rank test did show a significant correlation between the vertebral alignment angle induced by different head-neck positions and a normalized EDWI (ρ = 0.33, P < .0001). Student's t-test revealed that the presence of C6-C7 transposition of the transverse processes did not influence EDWI significantly. It was concluded that EDWI represents a reliable parameter for equine cervical radiographic intervertebral disc space width. Practical implementation of EDWI warrants monitoring in a group of adult horses while maintaining a standardized head-neck position.

Cervical articular process joint osteochondrosis in Warmblood foals.

BACKGROUND: In Warmblood horses, degenerative joint disease is involved in cervical malformation and malarticulation (CVM). The degree of contribution of articular process joint (APJ) osteochondrosis (OC) is not clear. OBJECTIVES: (a) To explore the presence of predilection sites for APJ OC in cervical and cranial thoracic vertebral columns of Warmblood foals and (b) to examine the correlation of such a site with the predilection site of CVM. STUDY DESIGN: Case series. METHODS: Seven hundred APJ facets of C2 to T2 of 29 foals (11 months gestation to 12 months [median age 7 days; range 365 days; 95% confidence interval [95% CI] 2-47 days]) were examined for OC and prevalence between joints, and the predilection site for CVM and the cranial cervical vertebral column were evaluated. RESULTS: About 20.6% of facets revealed OC. There was no predilection site. Prevalence decreased with age up to 1 year (odds ratio [OR] 0.997; (95% CI 0.975-0.998)) but not up to 5 months. Severity increased with age in all age ranges (up to 1 year OR 1.023; 95% CI 1.005-1.049; >1-5 months, OR 1.203; 95% CI 1.014e+00-1.921; up to 1 month, OR 1.114; 95% CI 1.041-1.228). Highest prevalence was in cranial facets of the cervical and cervical-thoracic joints and in caudal facets of the thoracic joint up to 1 year and up to 1 month (OR 0.364; 95% CI 0.170-0.745, OR 0.434; 95% CI: 0.235-0.782, OR 7.665; 95% CI: 1.615-66.553 and OR 0.400; 95% CI 0.170-0.880, OR 0.351; 95% CI 0.172-0.700, OR 5.317; 95% CI 1.098-44.344 respectively). MAIN LIMITATIONS: Two-thirds of the foals were less than 1 month of age. CONCLUSIONS: Articular process joint OC in Warmblood foals is common and is not more prevalent at CVM predilection sites, suggesting that abnormalities of enchondral ossification may not be major contributors to CVM.

Equine cervical intervertebral disc degeneration is associated with location and MRI features.

Morphology of the equine cervical intervertebral disc is different from that in humans and small companion animals and published imaging data are scarcely available. The objectives of this exploratory, methods comparison study were (a) to describe MRI features of macroscopically nondegenerated and degenerated intervertebral discs (b) to test associations between spinal location and macroscopic degeneration or MRI-detected annular protrusion and between MRI-detected annular protrusion and macroscopic degeneration, and (c) to define MRI sequences for characterizing equine cervical intervertebral disc degeneration. Ex vivo MRI of intervertebral discs was performed in 11 horses with clinical signs related to the cervical region prior to macroscopic assessment. Mixed-effect logistic regression modeling included spinal location, MRI-detected annular protrusion, and presence of macroscopic degeneration with "horse" as random effect. Odds ratio and 95% confidence interval were determined. Reduced signal intensity in proton density turbo SE represented intervertebral disc degeneration. Signal voids due to presence of gas and/or hemorrhage were seen in gradient echo sequences. Presence of macroscopic intervertebral disc degeneration was significantly associated with spinal location with odds being higher in the caudal (C5 to T1) versus cranial (C2 to C5) part of the cervical vertebral column. Intervertebral discs with MRI-detected annular protrusion grades 2-4 did have higher odds than with grade 1 to have macroscopic degeneration. It was concluded that MRI findings corresponded well with gross macroscopic data. Magnetic resonance imaging of the equine cervical intervertebral disc seems to be a promising technique, but its potential clinical value for live horses needs to be explored further in a larger and more diverse population of horses.

The Genomic Makeup of Nine Horse Populations Sampled in the Netherlands.

The spectrum of modern horse populations encompasses populations with a long history of development in isolation and relatively recently formed types. To increase our understanding of the evolutionary history and provide information on how to optimally conserve or improve these populations with varying development and background for the future, we analyzed genotype data of 184 horses from 9 Dutch or common horse populations in the Netherlands: The Belgian draft horse, Friesian horse, Shetland pony, Icelandic horse, Gelder horse, Groninger horse, harness horse, KWPN sport horse and the Lipizzaner horse population. Various parameters were estimated (e.g., runs of homozygosity and FST values) to gain insight into genetic diversity and relationships within and among these populations. The identified genomic makeup and quantified relationships did mostly conform to the development of these populations as well as past and current breeding practices. In general, populations that allow gene-flow showed less inbreeding and homozygosity. Also, recent bottlenecks (e.g., related to high selective pressure) caused a larger contribution of long ROHs to inbreeding. Maintaining genetic diversity through tailor-made breeding practices is crucial for a healthy continuation of the investigated, mostly inbred and (effectively) small sized horse populations, of which several already experience inbreeding related issues.

Sagittal plane fore hoof unevenness is associated with fore and hindlimb asymmetrical force vectors in the sagittal and frontal planes.

Asymmetry in forelimb dorsal hoof wall angles, termed unevenness, is associated with forelimb gait asymmetries, but compensatory mechanisms and out of plane ground reaction forces (GRFs) due to unevenness have yet to be documented. The aim of this study was therefore to investigate the effects of fore hoof unevenness on contralateral fore and hind limb force vectors patterns, in both sagittal and frontal planes. A group of n = 34 riding horses were classified into four groups: hoof angle difference of more than 1.5 degrees (UNEVEN; n = 27), including higher left fore (HIGH-LF; n = 12), higher right fore (HIGH-RF; n = 15), and hoof angle difference of less than 1.5 degrees (EVEN; n = 7). Three dimensional ground reaction forces GRFs were collected during trotting. GRF summary vectors representing the magnitude (VecMag) and angular direction (VecAng) of the entire stance phase in the sagittal and the frontal plane were calculated. The effects of unevenness on GRF production were explored using linear regression, repeated measures ANOVA and statistical parametric mapping (SPM) with significance at (P<0.05). In all uneven groups, increasing unevenness affected sagittal VecAng values in the forelimbs, with more propulsive GRF in the high hoof. In the HIGH-RF group, medial GRFs were also found in the high RF hoof compared to lateral GRFs in the low LF hoof (RF VecAng: 0.97±1.64 (deg); LF VecAng: -0.64±1.19 (deg); P<0.05). In both HIGH groups, compensatory associations to increasing unevenness were only found in the RH, but also a significantly greater lateral VecAng was found in the LH (P<0.05) compared to the RH limb. No significant differences (P>0.05) were found between hindlimb pairs in the EVEN group. Unbalanced sagittal and increased frontal plane GRFs in uneven horses suggest that they have greater locomotory challenges, as the equine musculoskeletal system is not constructed to withstand movement and loading in the frontal plane as effectively as it is in the sagittal plane.

EquiMoves: A Wireless Networked Inertial Measurement System for Objective Examination of Horse Gait.

In this paper, we describe and validate the EquiMoves system, which aims to support equine veterinarians in assessing lameness and gait performance in horses. The system works by capturing horse motion from up to eight synchronized wireless inertial measurement units. It can be used in various equine gait modes, and analyzes both upper-body and limb movements. The validation against an optical motion capture system is based on a Bland-Altman analysis that illustrates the agreement between the two systems. The sagittal kinematic results (protraction, retraction, and sagittal range of motion) show limits of agreement of ± 2.3 degrees and an absolute bias of 0.3 degrees in the worst case. The coronal kinematic results (adduction, abduction, and coronal range of motion) show limits of agreement of - 8.8 and 8.1 degrees, and an absolute bias of 0.4 degrees in the worst case. The worse coronal kinematic results are most likely caused by the optical system setup (depth perception difficulty and suboptimal marker placement). The upper-body symmetry results show no significant bias in the agreement between the two systems; in most cases, the agreement is within ±5 mm. On a trial-level basis, the limits of agreement for withers and sacrum are within ±2 mm, meaning that the system can properly quantify motion asymmetry. Overall, the bias for all symmetry-related results is less than 1 mm, which is important for reproducibility and further comparison to other systems.

Intervertebral Disc Degeneration in Warmblood Horses: Morphology, Grading, and Distribution of Lesions.

Equine intervertebral disc degeneration is thought to be rare and of limited clinical relevance, although research is lacking. To objectively assess pathological changes of the equine intervertebral disc and their clinical relevance, description of the normal morphology and a practical, biologically credible grading scheme are needed. The objectives of this study are to describe the gross and histological appearance of the equine intervertebral discs and to propose a grading scheme for macroscopic degeneration. Spinal units from 33 warmblood horses were grossly analyzed and scored. Of the 286 intervertebral discs analyzed, 107 (37%) were assigned grade 1 and grade 2 (considered normal) and were analyzed histologically. A nucleus pulposus and an annulus fibrosus could be identified macroscopically and histologically. Histologically, the nucleus pulposus was composed of a cartilaginous matrix and the annulus fibrosus of parallel collagenous bands. A transition zone was also histologically visible. Intra- and inter-observer reliability scores were high for all observers. Higher grades were associated with greater age. Gross changes associated with equine intervertebral disc degeneration (grades 3-5)-that is, yellow discoloration, cleft formation (tearing), and changes in consistency of the nucleus pulposus-were largely similar to those in humans and dogs and were most prevalent in the caudal cervical spine. Equine intervertebral disc degeneration was not associated with osteophyte formation. Changes of the vertebral bone were most common in the thoracolumbar spine but were not correlated with higher grades of intervertebral disc degeneration. Thus, changes of the vertebral bone should be excluded from grading for equine intervertebral disc degeneration.

Current insights into the molecular genetic basis of dwarfism in livestock.

Impairment of bone growth at a young age leads to dwarfism in adulthood. Dwarfism can be categorised as either proportionate, an overall size reduction without changes in body proportions, or disproportionate, a size reduction in one or more limbs, with changes in body proportions. Many forms of dwarfism are inherited and result from structural disruptions or disrupted signalling pathways. Hormonal disruptions are evident in Brooksville miniature Brahman cattle and Z-linked dwarfism in chickens, caused by mutations in GH1 and GHR. Furthermore, mutations in IHH are the underlying cause of creeper achondroplasia in chickens. Belgian blue cattle display proportionate dwarfism caused by a mutation in RNF11, while American Angus cattle dwarfism is caused by a mutation in PRKG2. Mutations in EVC2 are associated with dwarfism in Japanese brown cattle and Tyrolean grey cattle. Fleckvieh dwarfism is caused by mutations in the GON4L gene. Mutations in COL10A1 and COL2A1 cause dwarfism in pigs and Holstein cattle, both associated with structural disruptions, while several mutations in ACAN are associated with bulldog-type dwarfism in Dexter cattle and dwarfism in American miniature horses. In other equine breeds, such as Shetland ponies and Friesian horses, dwarfism is caused by mutations in SHOX and B4GALT7. In Texel sheep, chondrodysplasia is associated with a deletion in SLC13A1. This review discusses genes known to be involved in these and other forms of dwarfism in livestock.

A horse's locomotor signature: COP path determined by the individual limb.

INTRODUCTION: Ground reaction forces in sound horses with asymmetric hooves show systematic differences in the horizontal braking force and relative timing of break-over. The Center Of Pressure (COP) path quantifies the dynamic load distribution under the hoof in a moving horse. The objective was to test whether anatomical asymmetry, quantified by the difference in dorsal wall angle between the left and right forelimbs, correlates with asymmetry in the COP path between these limbs. In addition, repeatability of the COP path was investigated. METHODS: A larger group (n = 31) visually sound horses with various degree of dorsal hoof wall asymmetry trotted three times over a pressure mat. COP path was determined in a hoof-bound coordinate system. A relationship between correlations between left and right COP paths and degree of asymmetry was investigated. RESULTS: Using a hoof-bound coordinate system made the COP path highly repeatable and unique for each limb. The craniocaudal patterns are usually highly correlated between left and right, but the mediolateral patterns are not. Some patterns were found between COP path and dorsal wall angle but asymmetry in dorsal wall angle did not necessarily result in asymmetry in COP path and the same could be stated for symmetry. CONCLUSION: This method is a highly sensitive method to quantify the net result of the interaction between all of the forces and torques that occur in the limb and its inertial properties. We argue that changes in motor control, muscle force, inertial properties, kinematics and kinetics can potentially be picked up at an early stage using this method and could therefore be used as an early detection method for changes in the musculoskeletal apparatus.

Dwarfism with joint laxity in Friesian horses is associated with a splice site mutation in B4GALT7.

BACKGROUND: Inbreeding and population bottlenecks in the ancestry of Friesian horses has led to health issues such as dwarfism. The limbs of dwarfs are short and the ribs are protruding inwards at the costochondral junction, while the head and back appear normal. A striking feature of the condition is the flexor tendon laxity that leads to hyperextension of the fetlock joints. The growth plates of dwarfs display disorganized and thickened chondrocyte columns. The aim of this study was to identify the gene defect that causes the recessively inherited trait in Friesian horses to understand the disease process at the molecular level. RESULTS: We have localized the genetic cause of the dwarfism phenotype by a genome wide approach to a 3 Mb region on the p-arm of equine chromosome 14. The DNA of two dwarfs and one control Friesian horse was sequenced completely and we identified the missense mutation ECA14:g.4535550C > T that cosegregated with the phenotype in all Friesians analyzed. The mutation leads to the amino acid substitution p.(Arg17Lys) of xylosylprotein beta 1,4-galactosyltransferase 7 encoded by B4GALT7. The protein is one of the enzymes that synthesize the tetrasaccharide linker between protein and glycosaminoglycan moieties of proteoglycans of the extracellular matrix. The mutation not only affects a conserved arginine codon but also the last nucleotide of the first exon of the gene and we show that it impedes splicing of the primary transcript in cultured fibroblasts from a heterozygous horse. As a result, the level of B4GALT7 mRNA in fibroblasts from a dwarf is only 2 % compared to normal levels. Mutations in B4GALT7 in humans are associated with Ehlers-Danlos syndrome progeroid type 1 and Larsen of Reunion Island syndrome. Growth retardation and ligamentous laxity are common manifestations of these syndromes. CONCLUSIONS: We suggest that the identified mutation of equine B4GALT7 leads to the typical dwarfism phenotype in Friesian horses due to deficient splicing of transcripts of the gene. The mutated gene implicates the extracellular matrix in the regular organization of chrondrocyte columns of the growth plate. Conservation of individual amino acids may not be necessary at the protein level but instead may reflect underlying conservation of nucleotide sequence that are required for efficient splicing.

EX VIVO COMPUTED TOMOGRAPHIC EVALUATION OF MORPHOLOGY VARIATIONS IN EQUINE CERVICAL VERTEBRAE.

Diagnostic imaging is one of the pillars in the clinical workup of horses with clinical signs of cervical spinal disease. An improved awareness of morphologic variations in equine cervical vertebrae would be helpful for interpreting findings. The aim of this anatomic study was to describe CT variations in left-right symmetry and morphology of the cervical and cervicothoracic vertebrae in a sample of horses. Postmortem CT examinations of the cervical spine for horses without congenital growth disorders were prospectively and retrospectively recruited. A total of 78 horses (27 foals, 51 mature horses) were evaluated. Twenty-six horses (33.3%) had homologous changes in which a transposition of the caudal part of the transverse process (caudal ventral tubercle) of C6 toward the ventral aspect of the transverse process of C7 was present (n = 10 bilateral, n = 12 unilateral left-sided, n = 4 unilateral right-sided). There was one horse with occipito-atlantal malformation, two horses with rudimentary first ribs bilaterally, and one horse with bilateral transverse processes at Th1, representing homeotic (transitional) vertebral changes. Chi-square tests identified no significant differences in the number of conformational variations between the group of mature horses with or without clinical signs (P = 0.81) or between the group of mature horses and the group of foals (P = 0.72). Findings indicated that, in this sample of horses, the most frequently identified variations were homologous variations (transposition of the caudal part of the transverse process of C6-C7) in the caudal equine cervical vertebral column. Homeotic (transitional) variations at the cervicothoracic vertebral column were less common.

Musculoskeletal Disease in Aged Horses and Its Management.

Musculoskeletal disorders are the most prevalent health problem in aging horses. They are not life threatening, but are painful and an important welfare issue. Chronic joint disease (osteoarthritis) and chronic laminitis are the most prevalent. Treating osteoarthritis in the elderly horse is similar to treating performance horses, but aims at providing a stable situation with optimal comfort. Immediate medical treatment of flare-ups, long-term pain management, and adaptation of exercise and living conditions are the mainstays of treatment. Laminitis in the geriatric horse is related often to pituitary pars intermedia dysfunction, which may be treated with additional pergolide.

Clinicopathological findings in horses with a bi- or tripartite navicular bone.

BACKGROUND: Navicular bone partition is a rare condition reported in horses, which is during the evaluation of a lameness or prepurchase examination often misinterpreted for a parasagittal fracture. In this report, the clinicopathological findings of three cases of navicular bone partition are evaluated. The possible pathomechanisms underlying the condition are hypothesised, focusing on a potential origin of foetal vascular disturbance. This study is furthermore aiming at a clearer and earlier recognition of navicular bone partition, since this condition would finally predispose for a clinical lameness with a poor prognosis. CASE PRESENTATIONS: Case 1 was a 10-year-old Belgian Warmblood gelding with a Grade 3/5 chronic, recurrent left-forelimb lameness that had persisted for 4 months. Perineural palmar digital nerve block of the distal foot abolished the lameness. Radiographic examination revealed a bipartite navicular bone in the left forelimb. Unfortunately, the animal was lost to follow-up. Case 2 was a 7-year-old Quarter Horse stallion with a Grade 3/5 recurrent right forelimb lameness that had persisted for 2 years. The lameness switched to the contralateral left forelimb with a palmar digital nerve block. Radiographic examination identified a tripartite navicular bone in both forelimbs. Pathological examination additionally revealed chronic degenerative changes of the cartilage and subchondral bone with marked cystic changes. Case 3 was a 5-year-old Dutch Warmblood gelding with a Grade 3/5 recurrent left hindlimb lameness that had persisted for 6 months. Owing to the uncooperative behaviour of the horse, only a combined peroneal and tibial nerve block could be performed, which abolished the lameness. Radiographic examination revealed a bipartite navicular bone in the left hindlimb. Pathological examination showed a navicular bipartition in the left hindlimb, with microscopic changes comparable to those evident in Case 2; additionally, cartilage indentations were also found in the navicular bones of the right front- and hindlimb at a similar location as the partition site in the left hindlimb. CONCLUSIONS: It is speculated that a navicular bone partition has a congenital origin and is caused by vascular disturbance during foetal development. This may lead to aberrant endochondral ossification or the formation of multiple ossification centres resulting in navicular bone partitioning. In the adult horse, chronic repetitive biomechanical challenges at the partition sites may induce local degenerative changes with subchondral cyst formation and thus would cause a gradually developing chronic lameness with a poor prognosis.

Arthroscopic Removal of Large Extensor Process Fragments in 18 Friesian Horses: Long-Term Clinical Outcome and Radiological Follow-Up of the Distal Interphalangeal Joint.

OBJECTIVE: Report long-term clinical and radiological follow-up in horses after removal of large extensor process fragments occupying >25% of the joint surface of the distal interphalangeal joint (DIJ). STUDY DESIGN: Retrospective case series. ANIMALS: Friesian horses (n=18) that underwent arthroscopic removal of a large extensor process fragment. METHODS: Arthroscopic examination of the DIJ was performed in dorsal recumbency with the affected foot in extension using routine portals. Visualization of the fragment was improved using motorized synovial resectors. A dissection plane between the common digital extensor tendon and the extensor process fragment was created using sharp lever instruments, in some cases aided by motorized burrs and radiofrequency ligament dissection. The fragment was removed piecemeal using Ferris-Smith rongeurs. Medical records, preoperative and postoperative radiographs, and owner surveys were reviewed for case details and outcome. RESULTS: The technique described allowed removal of the large fragment in all 18 horses. Of the 17 horses where long-term clinical follow-up was available, 14 were used as intended and 3 kept some degree of lameness. The angle between the remodeled extensor process and the dorsal surface of the distal phalanx was increased and subchondral bone remodeling at the fragment bed was noted on postoperative lateromedial radiographs. CONCLUSION: Arthroscopic removal is a good treatment option for horses with large extensor process fragmentation with a good long-term outcome. Remodeling of the remaining extensor process and the subchondral new bone formation in the fragment bed can occur with functional recovery.

A longitudinal study on the performance of in vivo methods to determine the osteochondrotic status of young pigs.

BACKGROUND: In today's porcine industry, lameness has a major welfare and economic impact, and is often caused by osteochondrosis (OC). The etiological factors of the disease have been studied in depth, however, to this day, little is known about the natural course of the disorder and how it can be detected at an early stage in pigs. The aim of this pilot study was to assess the potential of three non-invasive techniques for the detection and monitoring of early OC processes in piglets. A group of weaned piglets (n = 19) were examined longitudinally using radiographs, a visual lameness scoring scheme and a quantitative pressure-mat based locomotion analysis system to detect OC in the humeroradial, femoropatellar and tarsocrural joints. At several time points, a selection of animals was euthanized for post-mortem examinations, including histology, which was the gold standard. RESULTS: In this study, clear signs of subclinical signs of OC were observed, however, we were unsuccessful in producing clinical OC. Lesions were observed to be commonly bilaterally symmetric in the joints examined in 80% of cases. The radiographic examinations showed a clear correlation with the gold standard, particularly when subclinical lesions were of a high histological score. Moreover, radiography was also able to detect the early repair processes, which appeared to take place at least until 14 weeks of age. Both visual scoring and pressure mat analyses showed good intra-assay reproducibility, with the pressure mat showing intra-class correlation values between 0.44 and 0.6 and the inter-observer agreement of visual scoring method was between 88 and 96%, however their correlation to OC lesions detected by histology was very weak, with only 2 out of 12 traits for the visual scoring method showing significant and biologically logical relations to a specific joint having histological OC lesions. For the pressure mat, only a maximum of 5 associations for specific joints with histological OC lesions were found out of a possible 8. CONCLUSION: All tested in-vivo methods showed good reproducibility. Radiography was the most reliable technique to detect and monitor longitudinally the earliest signs of OC in these piglets. It also demonstrated that the "Point of No Return" (PNR) of the disease, when repair processes end, might be later than anticipated, after 13 weeks of age. All in all, our study shows that the timing of the use of these in-vivo methods is critical to detect and monitor OC, especially in the early phases of the disease. It also shows the difficulty in producing OC regardless of the optimization of the experimental settings in relation to the etiological factors known to induce OC.

Clinical effects of buprenorphine on open field behaviour and gait symmetry in healthy and lame weaned piglets.

Lameness in pigs decreases animal welfare and economic profit for the farmer. An important reason for impaired welfare in lame animals is pain due to lameness. No direct measurement of pain is possible in animals, and methods to indirectly detect and quantify the amount of pain an animal is experiencing are urgently needed. In this study, two methods to assess pain associated with lameness in pigs were evaluated to determine if they were sensitive enough to detect a lameness reduction as an effect of an experimental analgesic medication. Asymmetry associated with lameness was objectively quantified using pressure mat kinetic parameters: peak vertical force (PVF), load rate (LR), vertical impulse (VI) and peak vertical pressure (PVP). Locomotor activity was assessed in an open field test. A dose of 0.04 mg/kg buprenorphine, a strong analgesic, was used to treat 10 lame pigs, while eight other lame pigs, treated with physiological saline solution, served as controls. Buprenorphine decreased lameness-associated asymmetry for pressure mat LR (P = 0.002), VI (P = 0.003) and PVP (P = 0.001) and increased activity of the lame pigs in the open field (P = 0.023), while saline-treated animals did not show any changes in asymmetry and became less active in the open field (P <0.001). It was concluded that measurement of gait asymmetry by pressure mat analysis and locomotor activity in an open field test are both sensitive enough to detect the analgesic effects of buprenorphine when used to treat moderate to severe clinical pain in a relatively small group of affected pigs. The methods used in this study may also provide promising additional tools for future research into early pain recognition and lameness treatment in pigs.

A nonsense mutation in B3GALNT2 is concordant with hydrocephalus in Friesian horses.

BACKGROUND: Hydrocephalus in Friesian horses is a developmental disorder that often results in stillbirth of affected foals and dystocia in dams. The occurrence is probably related to a founder effect and inbreeding in the population. The aim of our study was to find genomic associations, to investigate the mode of inheritance, to allow a DNA test for hydrocephalus in Friesian horses to be developed. In case of a monogenic inheritance we aimed to identify the causal mutation. RESULTS: A genome-wide association study of hydrocephalus in 13 cases and 69 controls using 29,720 SNPs indicated the involvement of a region on ECA1 (P <1.68 × 10(-6)). Next generation DNA sequence analysis of 4 cases and 6 controls of gene exons within the region revealed a mutation in ß-1,3-N-acetylgalactosaminyltransferase 2 (B3GALNT2) as the likely cause of hydrocephalus in Friesian horses. The nonsense mutation XM_001491545 c.1423C>T corresponding to XP_001491595 p.Gln475* was identical to a B3GALNT2 mutation identified in a human case of muscular dystrophy-dystroglycanopathy with hydrocephalus. All 16 available cases and none of the controls were homozygous for the mutation, and all 17 obligate carriers (= dams of cases) were heterozygous. A random sample of the Friesian horse population (n = 865) was tested for the mutation in a commercial laboratory. One-hundred and forty-seven horses were carrier and 718 horses were homozygous for the normal allele; the estimated allele frequency in the Friesian horse population is 0.085. CONCLUSIONS: Hydrocephalus in Friesian horses has an autosomal recessive mode of inheritance. A nonsense mutation XM_001491545 c.1423C>T corresponding to XP_001491595 p.Gln475* in B3GALNT2 (1:75,859,296-75,909,376) is concordant with hydrocephalus in Friesian horses. Application of a DNA test in the breeding programme will reduce the losses caused by hydrocephalus in the Friesian horse population.