Environmental factors of equine osteochondrosis and fetlock osteochondral fragments: A scoping review - Part 1.

Various environmental and genetic risk factors are linked to the pathogenesis of equine osteochondrosis and osteochondral fragments in the fetlock joint. Therefore, a scoping review was conducted to describe current evidence linking genetic factors and environmental factors of these osteochondral disorders. This article constitutes the first part of this scoping review and focuses on environmental factors, with the second part addressing genetic factors. To identify potentially relevant papers online bibliographical databases PubMed and Web of Science were utilised, supplemented with articles listed on the OMIA website (OMIA:000750-9796). After entry collection, removing duplicates, screening titles, abstracts, and full-text documents for eligibility, and manually searching reference lists of the remaining articles, a total of 212 studies was identified for this scoping review. First, an overview of the current understanding of the etiopathogenesis of equine osteochondrosis and osteochondral fragments in the fetlock joint is given. Subsequently, the article delves into the environmental factors associated with the prevalence of these disorders, which are categorized into foetal programming, biomechanical trauma and exercise, growth, anatomic conformation, nutrition, weaning, hormonal factors, bacterial infection, sex, date of birth, and other environmental factors. In conclusion, future research should adopt a multidisciplinary approach, emphasizing longitudinal studies and precise phenotype definitions. This strategy will help elucidate the complex relationships between environmental factors and OC, DOF, and POF, considering the dynamic nature, varying phenotypes, and scarcity of research in some domains of these osteochondral disorders. This approach will be crucial in developing effective management strategies aimed at improving equine orthopaedic health.

Association between navicular bone fragmentation and shape in Belgian Warmblood horses.

OBJECTIVES: To investigate whether in navicular bones (NB) from warmbloods, distal border fragmentation is associated with the shape of the proximal articular border or other radiological findings. METHODS: Radiographs of the front feet of 325 normal, subadult horses presented for admission as breeding stallions were reviewed. RESULTS: The proximal articular border of the total of 650 NB was classified as straight (n = 278), convex (n = 184), undulating (n = 147) or concave (n = 41). Distal border fragments were present in 57 NB (8.8%). They were significantly more prevalent in NB with a concave (9/41; 22%) or undulating (19/147; 13%) proximal articular border compared to NB with a straight (17/278; 6%) or convex shape (12/184; 7%). No other significant associations were found. CLINICAL SIGNIFICANCE: Hypothetically, since a shape-fragment association was found and the distribution of biomechanical forces exerted on the navicular bone are assumed to be shape dependent, distal border fragments may be a result of unfavourable loading of the navicular region.

Dynamic evaluation of toe-heel and medio-lateral load distribution and hoof landing patterns in sound, unshod Standardbred horses with toed-in, toed-out and normal hoof conformation.

Dynamic load distribution and landing patterns play an important role in equine orthopaedics. The aim of this study was to analyse dynamic load distribution and hoof landing patterns of sound Standardbreds. Twenty-four sound, unshod Standardbreds were walked and trotted over a pressure plate to obtain the peak vertical force (PVF), vertical impulse (VI) and stance time (ST) of both forelimbs. Asymmetry indices between both forelimbs were calculated and the dynamic medio-lateral and toe-heel load distribution of the forelimbs were compared between normal, toed-in and toed-out horses. The hoof landing patterns were determined on the first loaded zone. Linear mixed models and Chi-square analysis were used for statistical analysis (α = 0.05). At walk and trot, there were no significant differences in asymmetry of PVF, VI and ST between normal (n = 9/24 horses), toed-in (n = 6/24 horses), and toed-out horses (n = 9/24 horses). In both forelimbs, a flat landing (39.6% and 70.8% at walk and trot, respectively) predominated over all hoof conformations. No significant association between hoof conformation and landing pattern was observed (P > 0.05). Toed-out horses had significantly higher loading of the medial zone at the end of the stance phase at walk than normal horses (P = 0.002). No other statistically significant differences in load distribution were observed (P > 0.05). In conclusion, flat landing was more common in Standardbred horses than previously described in Warmbloods, emphasising the need for breed-specific clinical reference data and biomechanical studies. Moreover, mild deviations in hoof conformation did not necessarily affect load distribution.

The lipopolysaccharide model for the experimental induction of transient lameness and synovitis in Standardbred horses.

An established lipopolysaccharide (LPS) model previously described in Warmbloods, was inconsistent in Standardbred horses, where lameness was not detected despite the presence of synovitis. The present study aimed to determine the dose of LPS from E. coli O55:B5 required to induce mild to moderate lameness following middle carpal joint injection in Standardbred horses and to quantitate the induced lameness over time, with and without anti-inflammatory pre-treatment. In a baseline trial, eight healthy, clinically sound Standardbred horses were used in a rule-based dose-escalation design trial, starting at a dose of 10 endotoxin units (EU). Lameness at trot was evaluated visually and quantitatively (using an inertial-sensor system and pressure plate analysis). Synovial fluid aspirates were analysed for total nucleated cell counts, total protein and prostaglandin E2 (PGE2). Following 2 months wash-out, the effective LPS-dose determined in the baseline trial was used to evaluate the effect of anti-inflammatory treatment. A mixed model for repeated measures with horse as random effect was used for analysis. After injection of 10 EU LPS, the desired degree of lameness was observed in the baseline trial, with maximal lameness at post-injection hour (PIH) 4, followed by a rapid decline and return to baseline by PIH 48. No lameness was observed following pre-treatment with meloxicam. In synovial fluid, PGE2 was significantly higher at PIH 8 and PIH 24 in the baseline trial compared with following meloxicam pre-treatment. In conclusion, injection of the middle carpal joint with 10 EU LPS consistently induces a transient lameness and synovitis in Standardbred horses.

Use of blood culture medium enrichment for synovial fluid culture in horses: a comparison of different culture methods.

REASONS FOR PERFORMING STUDY: Standard methods for culturing equine synovial fluid (SF) are often unrewarding. Evidence-based information on the relative efficiency of different systems used for optimisation of isolation of microorganisms from equine SF is lacking. OBJECTIVES: To compare the results of different culture systems performed in parallel on SF samples from horses clinically diagnosed with synovial sepsis. METHODS: Synovial fluid specimens were collected between February 2007 and October 2008 from all horses admitted to a referral hospital that were clinically diagnosed with synovial sepsis and from control horses. Synovial fluid samples were cultured in parallel by: 1) direct agar culture (DA); agar culture after: 2) lysis-centrifugation pretreatment (LC); 3) conventional enrichment (CE); 4) combined LC/CE; or 5) blood culture medium enrichment using an automated system (BACTEC 9050). RESULTS: Ninety SF samples from 82 horses were included, together with 40 control samples. Seventy-one of 90 samples (79%) were culture-positive by using blood culture medium enrichment (BACTEC), which was significantly higher compared to all other methods. BACTEC enrichment was never negative while any of the other methods was positive. Although agar culture following LC and/or CE resulted in a slightly higher number of positive samples compared to DA, this difference was not significant. All control samples were culture negative by the 5 different techniques. Although the majority of samples containing isolates recovered without enrichment, culture results after BACTEC enrichment were available on the same day as for agar culture with or without LC (19/23 samples), while CE postponed recovery by at least one day in 20/23 samples. CONCLUSION: Blood culture medium enrichment is superior to other techniques for isolation of bacteria from SF of horses. The use of an automated system allows enrichment without substantially postponing recovery of microorganisms. POTENTIAL RELEVANCE: The efficient and fast isolation of microorganisms from infected SF by the BACTEC system allows for rapid susceptibility testing and a more appropriate antibiotic treatment.

Cardiorespiratory effects of enoximone in anaesthetised colic horses.

REASONS FOR PERFORMING STUDY: No studies have been reported on the effects of enoximone in anaesthetised colic horses. OBJECTIVE: To examine whether enoximone improves cardiovascular function and reduces dobutamine requirement in anaesthetised colic horses. METHODS: Forty-eight mature colic horses were enrolled in this prospective, randomised clinical trial. After sedation (xylazine 0.7 mg/kg bwt) and induction (midazolam 0.06 mg/kg bwt, ketamine 2.2 mg/kg bwt), anaesthesia was maintained with isoflurane in oxygen and a lidocaine constant rate infusion (15 mg/kg bwt, 2 mg/kg/h). Horses were ventilated (PaCO2 < 8.00 kPa). If hypotension occurred, dobutamine and/or colloids were administered. Ten minutes after skin incision, horses randomly received an i.v. bolus of enoximone (0.5 mg/kg bwt) or saline. Monitoring included respiratory and arterial blood gases, heart rate (HR), arterial pressure and cardiac index (CI). Systemic vascular resistance (SVR), stroke index (SI) and oxygen delivery index (DO2I) were calculated. For each variable, changes between baseline and T10 within each treatment group and/or colic type (small intestines, large intestines or mixed) were analysed and compared between treatments in a fixed effects model. Differences between treatments until T30 were investigated using a mixed model (a = 0.05). RESULTS: Ten minutes after enoximone treatment, CI (P = 0.0010), HR (P = 0.0033) and DO2I (P = 0.0007) were higher and SVR lower (P = 0.0043) than at baseline. The changes in CI, HR and SVR were significantly different from those after saline treatment. During the first 30 min after enoximone treatment, DO2I (P = 0.0224) and HR (P = 0.0003) were higher than after saline administration. Because the difference in HR between treatments was much clearer in large intestine colic cases, an interaction was detected between treatment and colic type in both analyses (P = 0.0076 and 0.0038, respectively). CONCLUSIONS: Enoximone produced significant, but short lasting, cardiovascular effects in colic horses. POTENTIAL RELEVANCE: Enoximone's cardiovascular effects in colic horses were of shorter duration than in healthy ponies.

Can the hoof be shod without limiting the heel movement? A comparative study between barefoot, shoeing with conventional shoes and a split-toe shoe.

Conventional shoeing restricts heel movement, which may have a negative effect on the orthopaedic health of the horse. A randomised crossover experimental study using noninvasive techniques was performed to compare the mediolateral heel movement in barefoot horses, horses shod with a conventional toe clipped shoe and with a new type of shoe with a split toe. In eight horses, 16 forelimbs were tested barefoot, shod with a conventional shoe and with the split-toe (ST) shoe, in random order. A displacement sensor was secured on the heels and measurements were collected continuously at a frequency of 679Hz while horses were exercised on a treadmill at the walk (1.8m/s), trot (3.5m/s) and canter (8m/s). Differences in heel movement between the conditions were analysed using a generalised estimating equations approach. The conventional shoe was associated with significantly less heel expansion compared with the ST shoe and barefoot situation in all gaits (P≤0.001). Heel expansion with the ST shoe was not significantly different from the barefoot condition. For all gaits, shoeing was associated with a significant reduction in heel contraction compared with the barefoot situation (P≤0.038), except for the heel contraction at the canter using a conventional shoe. In conclusion, the heel expansion with the ST shoe did not differ significantly from when the horse was barefoot, in contrast with the significant restriction of the heel movement when a conventional shoe was used.

The use of equine chondrogenic-induced mesenchymal stem cells as a treatment for osteoarthritis: A randomised, double-blinded, placebo-controlled proof-of-concept study.

BACKGROUND: There is a need to improve therapies for osteoarthritis in horses. OBJECTIVES: To assess the efficacy of equine allogeneic chondrogenic-induced mesenchymal stem cells combined with equine allogeneic plasma as a novel therapy for osteoarthritis in horses. STUDY DESIGN: Randomised, double-blinded, placebo-controlled experiment. METHODS: In 12 healthy horses, osteoarthritis was induced in the metacarpophalangeal joint using an osteochondral fragment-groove model. Five weeks after surgery, horses were randomly assigned to either an intra-articular injection with chondrogenic-induced mesenchymal stem cells + equine allogeneic plasma (= intervention) or with 0.9% saline solution (= control). From surgery until the study end, horses underwent a weekly joint and lameness assessment. Synovial fluid was collected for cytology and biomarker analysis before surgery and at Weeks 5, 5 + 1d, 7, 9 and 11. At Week 11, horses were subjected to euthanasia, and the metacarpophalangeal joints were evaluated macroscopically and histologically. RESULTS: No serious adverse events or suspected adverse drug reactions occurred during the study. A significant improvement in visual and objective lameness was seen with the intervention compared with the control. Synovial fluid displayed a significantly higher viscosity and a significantly lower glycosaminoglycan concentration in the intervention group. Other biomarkers or cytology parameters were not significantly different between the treatment groups. Significantly less wear lines and synovial hyperaemia were present in the intervention group. The amount of cartilage oligomeric matrix protein, collagen type II and glycosaminoglycans were significantly higher in the articular cartilage of the intervention group. MAIN LIMITATIONS: This study assessed the short-term effect of the intervention on a limited number of horses, using an osteoarthritis model. This study also included multiple statistical tests, increasing the risk of type 1 error. CONCLUSIONS: Equine allogeneic chondrogenic-induced mesenchymal stem cells combined with equine allogeneic plasma may be a promising treatment for osteoarthritis in horses. The Summary is available in Spanish - see Supporting Information.

Equine allogeneic chondrogenic induced mesenchymal stem cells: A GCP target animal safety and biodistribution study.

The safety of the intra-articular use of mesenchymal stem cells (MSCs) is scarcely reported. Therefore, the goal of this study was to investigate the safety of a single intra-articular injection with allogeneic chondrogenic induced MSCs combined with equine plasma (=the investigational product: IVP) compared to a saline (0.9% NaCl) placebo control (=control product: CP). Sixteen healthy experimental horses were randomly assigned to receive a single intra-articular injection with either the IVP (n=8) or the CP (n=8) in the left metacarpophalangeal joint. All horses underwent a daily clinical assessment throughout the entire study period of 42days to assess adverse events. Additionally, a local joint assessment and a lameness examination were performed daily during the first two weeks, and weekly the following 4weeks. Blood samples were taken weekly for hematological and biochemical analysis. At the end of the study period, horses of the IVP group were euthanized for a thorough necropsy and to check for biodistribution. Tissue samples of the injected joint were collected for histological examination. In both CP and IVP treated horses a mild transient subjective increase in periarticular temperature and lameness was noted after the intra-articular injection with no significant differences between the treatment groups. No distribution of the cells was found using immunohistochemistry and no ectopic tissue formation or signs of inflammation were found on histology. A single intra-articular injection of allogeneic chondrogenic induced MSCs combined with allogeneic plasma in horses had the same clinical side effects as an intra-articular injection with saline solution.

Upward fixation of the patella in the horse. A retrospective study.

The objective of the present study was to assess the incidence of upward fixation of the patella (UFP) in different types of patients and to evaluate the outcome of conservative and surgical treatment for correcting this condition. A particular attempt was made to find out whether corrective trimming and/or shoeing (CTS) is efficient in the conservative treatment of UFP, and whether a longer postoperative resting period reduces the risk of complications after medial patellar desmotomy (MPD). Medical records of 78 horses with intermittent or permanent UFP were analyzed retrospectively. Young animals and ponies were mostly affected, mainly during winter (P < 0.05). Seventy-six horses with UFP were treated conservatively, with follow-up being possible in 64 of them. This treatment, in which CTS seemed the most important aspect, was fully successful in 51.6% of these patients; 20.3% of them improved partially. In case of no response to conservative treatment, or in case of a permanent fixation, MPD was performed in 20 horses, which corrected UFP completely in 17 of the 18 followed-up patients. However, gait abnormalities were seen in seven of those 17 horses postsurgically, but with the incidence being lower in horses that had rested for at least three months (25%) compared to horses that had only rested for less than one month (66.6%). Results indicate that conservative treatment, with special attention for CTS, is worth trying before performing more radical procedures to correct UFP, and that a longer convalescence period after MPD is desirable.

Anatomical Description of the Presence and Variability of the Digital Manica Flexoria in the Equine Digital Flexor Tendon Sheath.

During endoscopy (tenoscopy) of the distal aspect of the equine digital flexor tendon sheath (DFTS), the digital manica flexoria can be visualized connecting the distal branches of the superficial digital flexor tendon. However, this structure has been inconsistently described and variably named in the veterinary literature. The objectives of this study were to describe the presence, configuration and variability of the digital manica flexoria in the equine distal limb. Dissection of 144 equine cadaveric limbs revealed the presence of this structure in all the feet, although different types and conformations were identified. In the forelimbs, a membranous digital manica flexoria predominated (94%; P < 0.001), in particular a synovial bridge type (83%; P < 0.001). In the hindlimbs, a tendinous digital manica flexoria predominated (93%; P < 0.001), in particular the oblique-crossing of tendinous bundles (61%; P < 0.001). Passage dorsal to the digital manica flexoria towards the distal DFTS was only possible in 22 of the 144 limbs, all forelimbs. Clinicians should be aware of the intra- and inter-individual anatomical variations of the digital manica flexoria to avoid misinterpretation during ultrasonographic and tenoscopic examinations of the DFTS.

Evaluation of direct Etest for antimicrobial susceptibility testing of bacteria isolated from synovial fluid of horses using enrichment bottles.

This study evaluated the Etest for direct antimicrobial susceptibility testing (AST) of bacteria from equine synovial specimens, incubated in BACTEC enrichment bottles. Ninety-four culture-positive broths were inoculated onto agar to directly determine the minimum inhibitory concentrations (MICs) of 13 antimicrobials, using the Etest (direct Etest). Results were compared with those obtained with the agar dilution reference method, the standard Etest, and the disc diffusion method, after subculture and standardisation of the inoculum. For categorical comparison of AST results, MICs were translated into susceptibility categories, using clinical breakpoints. The direct Etest predicted categorical susceptibility/resistance of bacteria from equine synovial fluid with acceptable accuracy (overall categorical agreement, 91%) and was more reliable than the disc diffusion test. The direct Etest was less accurate than the standard Etest for generating MICs ± 1 log dilution relative to the reference method (overall essential agreement, 69% vs. 89%). As the Etest generated a high percentage of inaccuracies with trimethoprim and sulfadiazine, these were less suitable antimicrobial agents for inclusion. In conclusion, the direct Etest reliably predicted the susceptibility of isolates from equine synovial fluid for the tested antimicrobials, except for trimethoprim and sulfadiazine. Since it did not require subculture and preparation of a standardised inoculum, direct Etest results were available at least 24 h earlier than with other methods, which could facilitate the diagnosis of synovial infections. However, when accuracy is prioritised over speed for MIC determination, the standard Etest is preferred over the direct Etest.

The preventive effects of two nutraceuticals on experimentally induced acute synovitis.

BACKGROUND: Nutraceuticals are often used in the management of equine osteoarthritis, but scientific evidence of their efficacy is lacking. OBJECTIVES: To study the preventive effects of two new nutraceuticals after the experimental induction of synovitis in comparison with positive and negative control treatments. STUDY DESIGN: Blinded, controlled, randomised experiment. METHODS: Twenty-four healthy Standardbred horses were randomly allocated to supplement AT (multi-ingredient, 28 days), supplement HP (collagen hydrolysate, 60 days), meloxicam (4 days) or placebo (60 days). Synovitis was induced in the right intercarpal joint by intra-articular injection of 0.5 ng lipopolysaccharide (LPS) of Escherichia coli while treatments were continued. Blood and synovial fluid were sampled before treatment, immediately prior to LPS injection, and at 8, 24 and 48 h post-injection. Synovial fluid samples were analysed for total nucleated cell count (TNCC), total protein (TP) and selected biomarkers (prostaglandin E2 [PGE2 ], interleukin-6 [IL-6], glycosaminoglycans [GAGs], type II collagen synthesis [CPII], matrix metalloproteinase [MMP]). Lameness was scored by visual examination and pressure plate analysis immediately prior to LPS injection, and at 8, 24 and 48 h post-injection. Clinical examinations were performed before treatment, immediately prior to LPS injection, at 2, 4 and 6 h post-injection, and then twice per day during the test period. RESULTS: Before treatment and intra-articular challenge, there were no statistically significant differences among the treatment groups for any of the parameters. After intra-articular challenge, the placebo group showed significantly higher synovial fluid TP, TNCC and PGE2 compared with the meloxicam group, although the model did not induce a relevant amount of lameness. Both nutraceuticals resulted in significantly lower synovial fluid TP, TNCC and PGE2 compared with placebo. No statistical differences in IL-6, GAGs, CPII or MMPs were observed among treatment groups. No adverse effects were observed. MAIN LIMITATIONS: Despite evidence of synovitis, lameness was too mild to detect. CONCLUSIONS: The preventive administration of these nutraceuticals showed anti-inflammatory effects in this validated synovitis model. Therefore, further studies of their clinical applicability are warranted.

The effect of perineural anaesthesia and handler position on limb loading and hoof balance of the vertical ground reaction force in sound horses.

REASONS FOR PERFORMING STUDY: The effects of handler position and perineural anaesthesia in sound horses need investigation to facilitate interpretation of pressure plate analysis in lame horses. OBJECTIVES: To evaluate the effect of handler position and perineural anaesthesia on limb loading and particularly hoof balance in sound horses. STUDY DESIGN: Nonrandomised crossover study. METHODS: Six sound horses were walked and trotted over a pressure plate, with the handler on the left and subsequently on the right side, and finally after a bilateral low palmar digital nerve block. One week later this procedure was repeated before and after a bilateral abaxial sesamoidean nerve block. Peak vertical force, vertical impulse and stance time of 5 hoof prints of both forelimbs were obtained (126 Hz), and toe-heel and mediolateral hoof balance curves of the vertical force were plotted throughout stance. Limb-loading and timing data and hoof balance data (beginning, middle and end of the stance phase) were statistically compared (handler left compared with right; before compared with after low palmar digital nerve block and abaxial sesamoidean nerve block; baseline at first measurement session compared with second). RESULTS: There were no significant effects of handler position and perineural anaesthesia on peak vertical force, vertical impulse and stance time. Hoof balance curves were not affected by handler position or perineural anaesthesia. The limb loading data and hoof balance curves were comparable for each horse over the one-week interval. CONCLUSIONS: Handler position and perineural anaesthesia do not have an effect on limb loading and toe-heel and mediolateral hoof balance in sound horses, which is of fundamental importance before embarking on pressure plate analysis for lameness diagnosis.

Diffusion of mepivacaine to adjacent synovial structures after intrasynovial analgesia of the digital flexor tendon sheath.

REASONS FOR PERFORMING STUDY: Controversy exists about the specificity of diagnostic analgesia of the digital flexor tendon sheath (DFTS) in horses. OBJECTIVES: To evaluate the degree of diffusion of mepivacaine from the equine DFTS to adjacent synovial structures. STUDY DESIGN: Crossover experiment. METHODS: Under general anaesthesia, the DFTS of one front and one hindlimb of 8 horses were injected simultaneously with mepivacaine. Synovial fluid samples of the injected DFTS, the adjacent metacarpo-/metatarsophalangeal (MCP/MTP) joint, proximal interphalangeal joint, distal interphalangeal joint, navicular bursa and contralateral MCP/MTP joint were collected 15 min post injection (T15) from one of the injected limbs and 60 min post injection (T60) from the other limb. Venous blood samples were obtained at T0, T15 and T60 to evaluate systemic distribution of mepivacaine. After a 2-week washout period, the procedure was repeated using the same limbs but reversing the time of sampling (front vs. hindlimbs). The concentration of mepivacaine in samples was measured with a commercial ELISA kit. RESULTS: Mepivacaine concentrations in the DFTS samples, at both T15 (5077 mg/l) and T60 (3503 mg/l), exceeded those estimated sufficient to produce synovial analgesia (100 mg/l or 300 mg/l). Mepivacaine was found in all synovial structures adjacent to the injected DFTS and in the contralateral MCP/MTP joints, but concentrations were low, with a maximum value of only 3.2 mg/l. With the exception of the navicular bursa samples, the mepivacaine concentrations in the adjacent synovial structures were significantly higher at T60 than at T15 (P<0.03). Significantly higher mepivacaine concentrations were found in the ipsilateral than the contralateral MCP/MTP joints at T60 (P<0.001). Blood samples had significantly higher mepivacaine concentrations at T15 and T60 than at T0 (P<0.001). CONCLUSIONS: Mepivacaine injected into the DFTS of horses diffuses towards adjacent synovial structures without achieving clinically relevant concentrations.

Recording of cord dorsum potentials in calves.

Cord dorsum potentials are sensory evoked potentials being used to assess proximal sensory nerve, dorsal nerve root and spinal cord dorsal horn function. The purpose of the present study was to establish normal values for onset latency, peak latency and peak-to-peak amplitude of cord dorsum potentials after saphenous nerve stimulation in 15 healthy calves. Under general anesthesia, the saphenous nerve was stimulated in the periphery and cord dorsum potentials were recorded from the interarcuate space L3-L4, L4-L5, L5-L6 and L6-S1. Cord dorsum potentials were easily recorded at the different recording sites and consisted of a large negative peak followed by a long latency positive phase. The largest responses were recorded at the L5-L6 interarcuate space. Body temperature significantly influenced peak-to-peak amplitude. Onset latency prolonged with increasing limb length.

Detection of bacterial DNA in synovial fluid from horses with infectious synovitis.

Standard culturing techniques are often unrewarding in confirming diagnosis of synovial infection in the equine patient. Several human studies report the use of sensitive polymerase chain reaction (PCR) techniques for the detection of bacterial involvement in acute synovitis. However, successful extraction of bacterial DNA directly from clinical samples from horses without prior culture has not been reported yet. The goal of this study was to develop a sensitive and reliable method for molecular detection and identification of bacterial species in synovial fluid from horses with infectious synovitis. Synovial fluid samples from 6 horses with culture confirmed synovial infection were used for broad range 16S rRNA gene PCR. Synovial aspirates of 2 healthy horses were used as negative controls. Following extraction and purification of synovial fluid DNA, all samples were processed by touchdown PCR. Amplicons were detected by reverse line blot hybridisation and visualised with chemiluminescence. Pathogen-specific detection of 16S rRNA gene sequences was successful in all 6 synovial fluid samples. No bacterial DNA was detected in the aspirates from the negative control horses using touchdown PCR followed by 25 additional cycles of amplification. The identity of the pathogens was confirmed by DNA sequencing of the amplicons. It can be concluded that broad range 16S rRNA gene PCR followed by reverse line blot hybridisation is a promising technique for detection of bacterial DNA in synovial fluid samples. Further research should aim at the detection of bacterial DNA in synovial fluid samples suspected of infection but having negative culture results. When the 16S PCR proves to be reliable and more sensitive than standard culturing techniques, it may become a powerful tool in the diagnosis of synovial infection.

Evaluation of excision, cryosurgery and local BCG vaccination for the treatment of equine sarcoids.

Ninety-five horses with sarcoids were subjected to three types of treatment: surgical excision (conventional or carbon dioxide laser), cryotherapy or local BCG vaccination. The type of treatment was selected on the basis of the size, location and clinical appearance of the tumours. The choice between conventional and laser excision was empirical. A successful outcome was obtained in 11 of 14 (79 per cent) of the horses treated by cryosurgery, 18 of 27 (67 per cent) treated by BCG vaccination, 18 of 22 (82 per cent) treated by conventional excision, and 20 of 28 (71 per cent) treated with a carbon dioxide laser. For both excision methods, rigorous measures were taken to avoid autoinoculation and to ensure a wide margin of normal skin. The probability of local recurrence after excision was significantly higher for large sarcoids and sarcoids which had previously failed to respond to treatment. In 10 of the 31 horses with remaining sarcoids, some or all of the untreated sarcoids were observed to regress spontaneously.

A preliminary study on pressure-plate evaluation of forelimb toe-heel and mediolateral hoof balance on a hard vs. a soft surface in sound ponies at the walk and trot.

REASONS FOR PERFORMING STUDY: Thus far, pressure-plate analysis has been limited to measurements on a hard surface, whereas equine athletes routinely perform on a deformable substrate. OBJECTIVES: To explore pressure-plate analysis on arena footing. STUDY DESIGN: Nonrandomised crossover study using noninvasive techniques. METHODS: Five sound, unshod ponies were walked and trotted over a pressure plate in 2 different conditions. In the first session, the plate was covered with only a 5 mm rubber mat, and in the second session, with an additional 50 mm layer of sand and synthetic fibres. Limb loading and timing variables (peak vertical force, vertical impulse, peak vertical pressure, hoof contact area and stance phase duration) were obtained. Toe-heel and mediolateral hoof balance of the vertical force were calculated throughout stance (126 Hz). RESULTS: Peak vertical force, vertical impulse, peak vertical pressure and stance phase duration were decreased on the soft substrate, while hoof contact area increased. At impact, there was more even load distribution between the toe and heel region on the soft vs. hard surface. At mid-stance, there was more loading of the toe region on the soft compared with the hard surface. At impact, there was more even load distribution between lateral and medial on the soft vs. hard surface at walk, but not at trot. CONCLUSIONS: Comparison of observed vs. expected impulse indicated that the main factor involved in the lower loading on the soft surface is the dampening effect of the soft substrate, although definitive conclusions require concurrent force-plate analysis. The pressure plate permitted quantitative evaluation of hoof balance of the vertical force on a deformable surface at the walk and trot and proved that there is more even load distribution on arena footing.

Distal limb desensitisation following analgesia of the digital flexor tendon sheath in horses using four different techniques.

REASONS FOR PERFORMING STUDY: Controversy exists about the desensitisation obtained after diagnostic analgesia of the digital flexor tendon sheath (DFTS) during lameness examinations. OBJECTIVES: To determine whether DFTS analgesia results in inadvertent desensitisation of the palmar/plantar digital nerves and whether this depends on the injection technique used. STUDY DESIGN: Crossover experimental study. METHODS: The DFTS of 9 horses were injected with local anaesthetic solution and radiodense contrast medium using one of the following techniques: Proximal (at lateral proximal recess of the DFTS), Axial (axial to the lateral proximal sesamoid bone), Base (at base of the lateral proximal sesamoid bone), and Distal (at palmar/plantar mid-pastern). In total, 72 injections were performed. Skin desensitisation at the heel bulbs was tested with a dynamometer before and at 15, 30, 90 and 120 min after injection. RESULTS: Overall, complete desensitisation of a heel bulb at one or more time points after injection occurred in 22 limbs (30.6%). An additional 7 limbs were partially desensitised. Complete skin desensitisation occurred in 10, 3, 4 and 5 limbs using the Proximal, Axial, Base and Distal techniques respectively. Significant differences between techniques were only found at T30. The probability of skin desensitisation at the heel bulbs was 4 times higher when using the Proximal compared with the Axial and Base techniques in the forelimbs, and 3 times higher compared with the Axial and Distal techniques in the hindlimbs. Skin desensitisation nearly always occurred exclusively on the lateral heel bulb. Bilateral desensitisation only occurred in 5 limbs. CONCLUSIONS: Anaesthesia of the palmar/plantar digital nerves with distal limb desensitisation often occurs after DFTS analgesia. A higher chance of desensitisation exists when injecting the proximal DFTS recess. It is advisable to verify skin sensitivity at the heel bulbs after DFTS analgesia to avoid false interpretations about the origin of pain causing lameness.