Willem Back

An ELISA for detection of antibodies against influenza A nucleoprotein in humans and various animal species.

SummaryA double antibody sandwich blocking ELISA, using a monoclonal antibody (MAb) against influenza A nucleoprotein (NP) was developed to detect antibodies against influenza. Collections of serum samples were obtained from human and various animal species. All influenza A subtypes induced antibodies against hemagglutinins and NP. A close correlation between titers of the hemagglutination inhibition (HI) test and the NP-ELISA was seen. Antibodies against influenza NP were demonstrated in serum samples from humans, ferrets, swine, horses, chickens, ducks, guinea pigs, mice, and seals. The serum samples were collected at intervals during prospective epidemiological studies, from experimental and natural infections, and vaccination studies. The decline of maternal antibodies was studied in swine and horses. The NP-ELISA enables rapid serological diagnosis and is suited for influenza A antibody screening, especially in species which harbor several influenza subtypes. The HI and neuraminidase inhibition tests, however, must still be used for subtyping.

Dynamic pressure measurements for the detailed study of hoof balance: the effect of trimming

REASONS FOR PERFORMING STUDY Studies on hoof balance have, so far, only assessed the effects of strong and relatively unrealistic interventions due to technical limitations of measuring equipment. For the assessment of more subtle interventions, other techniques are necessary. OBJECTIVES To test a sensitive pressure measurement system during locomotion and to set a standard for further studies by using the system to evaluate the effects of trimming. METHODS Eighteen horses were measured before and after trimming with an interval of 4 weeks. Trimming was standardised to a straight hoof-pastern axis. The horses trotted over a pressure/force measuring system, with a temporal resolution of 240 Hz and a spatial resolution of 0.39 cm2. RESULTS The preferred way of landing was lateral, asymmetrical in both front and hind feet. The duration of landing was shorter in forelimbs than in hindlimbs with an equal decreasing trimming effect. Horses had a fixed hoof-unrollment pattern; the centre of pressure (CoP) travelled towards a maximum lateral deviation and returned towards the dorsopalmar/plantar axis of the hoof. Trimming decreased the intra-individual left/right difference in maximum lateral displacement. CONCLUSIONS The technique used provided easily accurate data to quantify hoof balance characteristics and to measure short-term trimming effects. POTENTIAL RELEVANCE Determination of CoP patterns has added value in gait analysis and can improve our understanding of the effects of different interventions on hoof balance.

Kinematic gait analysis in equine carpal lameness.

Gait analysis plays a major role in the clinical evaluation of equine lameness. It is generally accepted that the clinician expresses the grade of lameness as a subjective score. In this study lameness was objectively assessed using a standardized transient lameness model, in which lameness was induced by intra-articular injection of bacterial endotoxin into the radiocarpal joint of ponies. Lameness was scored by an experienced clinician, and locomotion was recorded simultaneously using a CODA-3 apparatus. The obtained kinematic gait parameters correlated well with the clinical lameness score and also provided possibilities to objectively study the locomotor disturbances of the lame limb in more detail at the walk and trot.

The role of electromyography in clinical diagnosis of neuromuscular locomotor problems in the horse.

REASONS FOR PERFORMING STUDY Systematically performed EMG needle examination of muscles provides essential information about the functional aspects of the motor unit. However, clinical studies in which information is given on the diagnostic and discriminative values of electromyography (EMG) in the horse are scarce. OBJECTIVES To determine to what extent inclusion of EMG analysis in clinical examination contributes to determination of type and localisation of abnormality. METHODS EMG analysis, complete clinical examination and diagnosis of 108 horses (mean +/- s.d. age 75 +/- 3.8 years; bodyweight 548 +/- 86 kg; height 1.67 +/- 0.07 m) were performed, and results without and with EMG analysis compared. RESULTS Without EMG, myopathy and neuropathy were diagnosed in 20 and 58 horses, respectively, and with EMG in 17 and 82 horses. EMG changed localisation in myopathy and neuropathy in 12 and 37% of cases, respectively. Lesions in the C1-T2, T2-L3 and L3-S3 segments were, respectively, diagnosed without EMG in 7, 11 and 30%, and with EMG in 27, 7 and 17% of cases. Where no clinical diagnosis could be made prior to EMG, many patients appeared to be suffering from localised cervical lesions (29%) or generalised neuropathy (54%). CONCLUSIONS AND POTENTIAL RELEVANCE The assistance of EMG in discriminating between normal, neuropathy and myopathy, and in locating pathology, contributes to diagnosis of neuromuscular problems.

Kinematic detection of superior gait quality in young trotting warmbloods

This study was conducted to identify objective criteria to select young horses with a good gait, which is a prerequisite for good performance in adult horses. The trot of 24 26-month-old Dutch Warmbloods, led on a loose shank, was subjectively scored by a judge and objectively assessed on a treadmill by using kinematic analysis equipment. It appeared that forelimb and hind limb stride and swing duration, scapula rotation, forelimb maximal fetlock extension, forelimb maximal retraction, hind limb maximal protraction, maximal stifle flexion, and maximal tarsal flexion significantly correlated with a generally accepted gait score in which length, suppleness, and strength are judged. Moreover, the ranking of the individual horses on the basis of gait quality according to their objectively measured kinematic variables was similar to the subjective ranking given by the judge. Thus, a complete picture was obtained of the variables in equine locomotion that determine the quality of the trot in warmbloods.

Intra-articular opioid analgesia is effective in reducing pain and inflammation in an equine LPS induced synovitis model.

REASONS FOR PERFORMING STUDY Intra-articular administration of morphine as a local analgesic and anti-inflammatory drug is widely used in human medicine. In equids, little is known about its clinical analgesic and anti-inflammatory efficacy. OBJECTIVES To use an inflammatory orthopaedic pain model to investigate the analgesic and anti-inflammatory effects of intra-articularly administered morphine as a new treatment modality in horses with acute arthritis. METHODS In a crossover study design, synovitis was induced in the left or right talocrural joint by means of intra-articular injection of 0.5 ng lipopolyssacharide (LPS). The effect of 120 mg morphine, intra-articularly administered at 1 h after induction of synovitis, was evaluated using both physiological and behavioural pain variables. Synovial fluid was sampled at 0, 4, 8, 28 and 52 h after induction of synovitis and analysed for total protein concentration, leucocyte count and for prostaglandin E(2), bradykinin and substance P concentrations by ELISA. Ranges of motion of metatarsophalangeal and talocrural joints were measured as kinematic variables with the horses walking and trotting on a treadmill under sound and lame conditions. Clinical lameness scores and several behavioural variables related to the perception of pain were obtained. RESULTS LPS injection caused marked transient synovitis, resulting in increased concentrations of inflammatory synovial fluid markers, clinical lameness, joint effusion and several behavioural changes, such as increased time spent recumbent, decreased limb loading at rest and decreased time spent eating silage. Intra-articular morphine resulted in a significant decrease in synovial white blood cell count, prostaglandin E(2) and bradykinin levels and improvement in clinical lameness, kinematic and behavioural parameters, compared to placebo treatment. CONCLUSIONS Intra-articular morphine offers potent analgesic and anti-inflammatory effects in horses suffering from acute synovitis. POTENTIAL RELEVANCE Local administration of opioids may be useful for horses with acute inflammatory joint pain and offers possibilities for multimodal analgesic therapies without opioid-related systemic side effects.

Uneven feet in a foal may develop as a consequence of lateral grazing behaviour induced by conformational traits

REASONS FOR PERFORMING STUDY Conformational traits are important in breeding, since they may be indicative for performance ability and susceptibility to injuries. OBJECTIVES To study whether certain desired conformational traits of foals are related to lateralised behaviour while foraging and to the development of uneven feet. METHODS Twenty-four Warmblood foals, born and raised at the same location, were studied for a year. Foraging behaviour was observed by means of weekly 10 min scan-sampling for 8 h. A preference test (PT) was developed to serve as a standardised tool to determine laterality. The foals were evaluated at age 3, 15, 27 and 55 weeks. The PT and distal limb conformation were used to study the relation between overall body conformation, laterality and the development of uneven feet. Pressure measurements were used to determine the loading patterns under the feet. RESULTS About 50% of the foals developed a significant preference to protract the same limb systematically while grazing, which resulted in uneven feet and subsequently uneven loading patterns. Foals with relatively long limbs and small heads were predisposed to develop laterality and, consequently unevenness. CONCLUSIONS Conformational traits may stimulate the development of laterality and therefore indirectly cause uneven feet.

Hoof growth between two shoeing sessions leads to a substantial increase of the moment about the distal, but not the proximal, interphalangeal joint

REASONS FOR PERFORMING STUDY There is little insight into the effects of routine farriery on the internal structures of the distal limb in sound horses. OBJECTIVES To measure the effect of change in hoof conformation during a shoeing interval on the moments about the proximal and distal interphalangeal joints (PIPJ, DIPJ) and to determine whether and how the horse compensates for this change in hoof conformation. METHODS Both front feet of 9 sound Warmblood horses were measured while standing on a pressure-force measuring system and radiographed in a lateromedial direction shortly after shoeing and 8 weeks later. From these data, ground reaction forces (GRF) and lever arms were measured in order to calculate joint moments. RESULTS After 8 weeks, the moment about the PIPJ did not increase significantly, but the moment about the DIPJ did so, indicating a compensatory mechanism for a change in hoof conformation in the DIPJ. CONCLUSIONS Standing horses compensate for hoof conformation change during an 8-week shoeing interval, which leads to increased DIPJ extension and consequently an increased loading of the deep digital flexor tendon. POTENTIAL RELEVANCE This study quantifies the effect of a shoeing interval on the internal structures of the foot and helps to determine an appropriate shoeing interval for individual horses in which the hoof with the lowest hoof angle is the best indicator. The exact determination of an optimal individual shoeing interval requires further study.

Quantification of the locomotion of Dutch warmblood foals

To get an in-depth insight into the ontogeny of the equine locomotor apparatus, the locomotion of 28 Dutch Warmblood foals was recorded at the walk, trot and canter on a treadmill using a CODA-3 apparatus. A great similarity in joint angle-time diagrams was found, although considerable differences in kinematic gait parameters of individual foals could be detected. These results contribute to an objective comparison of the locomotor performance of foals.

Genome-wide SNP association-based localization of a dwarfism gene in Friesian dwarf horses.

The recent completion of the horse genome and commercial availability of an equine SNP genotyping array has facilitated the mapping of disease genes. We report putative localization of the gene responsible for dwarfism, a trait in Friesian horses that is thought to have a recessive mode of inheritance, to a 2-MB region of chromosome 14 using just 10 affected animals and 10 controls. We successfully genotyped 34,429 SNPs that were tested for association with dwarfism using chi-square tests. The most significant SNP in our study, BIEC2-239376 (P(2df)=4.54 × 10(-5), P(rec)=7.74 × 10(-6)), is located close to a gene implicated in human dwarfism. Fine-mapping and resequencing analyses did not aid in further localization of the causative variant, and replication of our findings in independent sample sets will be necessary to confirm these results.